La domestication des plantes a débuté il y a quelques milliers d’années quand les hommes se sont sédentarisés. Ils ont sélectionné les plantes sauvages portant des caractères phénotypiques d’intérêt pour la consommation et production humaine. Ce processus évolutif a par conséquent modifié le patrimoine génétique des espèces domestiquées. Cette thèse se penche sur les traces génétiques induites par la domestication chez trois espèces de Solanacées : l’aubergine (Solanummelongena), le piment (Capsicum annuum) et la tomate (S. lycopersicum). En effet, si les caractères phénotypiques des plantes cultivées ont été sélectionnés depuis des milliers d’années, les conséquences moléculaires d’une telle sélection restent peu étudiées à l'échelle du génome. Cette étude est basée sur des données de diversité et d’expression de gènes (RNAseq). En utilisant des méthodes comparatives entre des variétés cultivées et leurs espèces sauvages apparentées, j’a iétudié, à l’échelle intra-spécifique, d’une part les histoires démographiques de chacune des espèces,et d’autre part les changements de diversité nucléotidique et d’expression des gènes dus à la domestication. La comparaison de ces trois événements indépendants de domestication, offre l’opportunité de décrypter les changements génétiques qui convergent chez ces trois espèces lors du processus de sélection humaine.Suite à une introduction qui pose le cadre de cette étude et présente l’état de l’art, le premier chapitre, s’inscrit dans un ouvrage portant sur la génomique des populations d’espèces modèles. Il propose une synthèse des connaissances accumulées en plus d’un siècle de recherche sur l’espèce modèle qu’est la tomate (S. lycopersicum). Ce chapitre permet également de compléter le contexte scientifique dans lequel cette thèse s’inscrit, notamment, en retraçant l’importance que les espèces sauvages apparentées ont eu dans l’amélioration de l’adaptabilité des variétés cultivées actuelles.L’hypothèse du deuxième chapitre révèle la convergence des changements démographiques entre les trois espèces malgré leurs événements indépendants de domestication. L’étude comparée d’inférences de scénarios démographiques a permis de reconstruire l’histoire démographique de chaque espèce cultivée. Ces inférences ont aussi facilité l’estimation des paramètres tels que les flux migratoires entre les espèces sauvages et cultivées, la force des goulots d’étranglement liés à l’intensité de la sélection humaine et la durée des événements de domestication. Ce chapitre permet de démontrer que les changements démographiques liés à la domestication dépendent de l’état de sympatrie ou d’allopatrie des variétés cultivées avec leurs sauvages apparentées. Les connaissances quant à la datation des événements de domestication de nos trois espèces restent très faibles, et les inférences ont permis d’établir des estimations de durée de domestication relativement précise. Ces nouvelles connaissances apportent une plus-value à cette étude pour nos trois espèces et nous invitent à s’interroger sur les différents compartiments du génome qui ont été sélectionnées et modifiées lors de la domestication.Le troisième chapitre teste l’hypothèse d’une convergence évolutive des changements moléculaires, notamment transcriptionnels, induits par la domestication et l’amélioration moderne.La comparaison des variétés cultivées à leurs espèces sauvages apparentées permet d’évaluer la convergence des mécanismes de régulation et d’adaptation liés à la domestication. C’est en testant la corrélation entre les traces génétiques (diversité nucléotidique) de sélection et les changements d’expression des gènes observés chez les variétés cultivées que l’hypothèse de départ a été validée.Cette analyse montre que la domestication, au-delà même de changements nucléotidiques, a modifié l’expression des gènes chez les trois espèces. / Domestication started thousand years ago when human shifted from hunter-gatherer to agrarian societies. They started selecting wild plants for phenotypes related to consumption andyield. This evolutionary process induced changes in the gene pool of domesticated plants. This thesis focuses on genetic footprints induced by domestication within a trio of Solanaceae species: the eggplant (Solanum melongena), the pepper (Capsicum annuum) and the tomato (S. lycopersicum).Crop plants have been selected for thousand years on phenotypic traits, but the molecularconsequences of such selection remain unknown at the genome-wide scale. The study was performed on a RNAseq data set; using comparative methods between crops and their wild relatives,I studied, at the intra-specific scale, the demographic history, and, both the nucleotide diversity and the gene expression changes due to domestication. Comparing these three independent events ofdomestication, is a great opportunity to decipher the interspecific genetic changes, converging for the three species, during the human selection process.The first chapter is a book chapter about population genomics in model species. It details thestate of art of hundred years of research on tomato as model species (S. lycopersicum). Tomato is amodel species in genetics, as well as in population genomics thanks to the important collection of genomic data that have been accumulating over years. Tomato has the strongest economic importance within the trio of studied species. By highlighting the importance of crop wild relative species for adaptability improvement of modern cultivars, this chapter describes the scientific context of this thesis work.The two next chapters are following these researches and show the importance to both conserve and study the crop wild relative species.In the second chapter, I hypothesize that demographic changes within the three species experience a convergence, despite their independent domestication events. The comparative studyof demographic inferences allows the reconstruction of each domesticated species demographichistory. Theses inferences facilitate the parameter estimations such as the migration rate between crop and wild, the bottleneck strength paired with the human selection and the duration of thedomestication events. This chapter reveals a common bottleneck phenomenon as well as migration rate dependent to the allopatric or sympatric state of the crops with their wild relatives. Knowledge concerning the domestication events dating, for each of the three species, remain poorly studied and this thesis work discloses relative domestication time durations.These new insights bring valuable knowledge to the three species and induce a questioning on thedifferent genome parts that are selected and modified through domestication.The third chapter, test the hypothesis of a convergent evolution of molecular changes,especially transcriptional, induced by domestication and modern breeding. The comparative analysis of crop plants and their wild relatives assesses the convergence of regulation and adaptation mechanisms due to domestication. By testing the correlation between the selection footprints on genes and the gene expression changes in crop compared to their wild relative species, the previous hypothesis was confirmed. This analysis implies that domestication modified gene expression in the three species beyond only nucleotide polymorphisms. The ortholog analysis of our species genes, confirmed that domestication facilitated the fruit development and plant growth but relaxed selective pressure on genes of plant defense and environmental stresses tolerance. Demonstrating demographic changes and molecular footprints of domestication, my PhD thesis highlights several proofs of convergence.

As bases moleculares da diversidade fenotipica dentro de espécies são de grande interesse aos biólogos evolutivos porque a evolução morfológica adaptival depende da seleção de variantes genéticas. Entretanto, há poucos exemplos da variação fenotípica cuja a base molecular é compreendida, especialmente entre animais vertebrados. Biólogos em geral concordam que o processo da seleção natural é a fonte predominante de diversificação morfológica. Tem sido documentado previamente que existe diversificação adaptativa da morfologia craniana entre os taxa mais elevados de Platyrrhini e também em espécies e em muitos géneros, que aparentemente diversificaram na morfologia do crânio por seleção natural. O hormônio de crescimento (GH) é um hormônio multifunctional, produzido principalmente pela glândula pituitária de todos os animais vertebrados para regular o metabolismo e para promover o crescimento pos-natal linear. A evolução molecular do GH foi estudada extensivamente em um grande número espécies de vertebrados, incluindo primatas. A evolução rápida do gene do GH em primatas, especiamente em regiões funcionalmente importantes, sugere a seleção darwiniana positiva. Entretanto, o relaxamento da seleção purificadora depois de múltiplas duplicações do locus GH não podem ser descartadas. O objetivo deste estudo era investigar a evolução molecular do gene do GH em primatas neotropicais, tentando identificar diferenças potenciais e funcionais entre taxa. As análises de máxima verossimilhança deste trabalho mostraram que a relação dN/dS no gene GH1 de primatas de Neotropicais é diferente entre gêneros, demonstrando que a evolução do GH1 no Platyrrhini não é compatível com o modelo neutro da evolução molecular. Estes resultados sugerem que o gene GH1 está sobre seleção positiva em alguns sítios na proteína durante o processo de diversificação dos primatas de Neotropical. / The molecular bases of phenotipic diversity within and between species are of great interest to evolutionary biologists because the adaptive morphological evolution depends on the selection of genetics variants. However, there are few examples of phenotypic variation whose molecular basis is understood, especially among vertebrates, while most biologists agree that the natural selection process is the predominant source of morphological diversification. Is has been previously documented that there is adaptive diversification of cranial morphology among the higher taxa of Platyrrhini and also in species in most genera that apparently diversified in skull morphology by natural selection. The Growth Hormone (GH) is a multifunctional hormone, mainly produced by the pituitary gland of all vertebrates to modulate the metabolism and promote linear postnatal growth. The molecular evolution of GH has been extensively studied in a large number of vertebrate species, including primates. The rapid GH gene evolution on primates, especially on sites that are functionally important, suggest positive Darwinian selection. However, the hypothesis of purifying selection relaxation after multiple duplications of the GH locus cannot be ruled out either. The aim of this study was to investigate the molecular evolution of the GH gene on neotropical primates, trying to identify potential and functional differences between taxa. The analyses of Maximum likelihood of this work has shown that the ratio dN/dS in the GH1 gene of Neotropical primates are different among genera, demonstrating that the evolution of the GH1 in the Platyrrhini is not compatible with the neutral model of molecular evolution. These results suggest that the GH1 gene suffered positive selection at some sites inside the protein during the process of diversification of the Neotropical primates.

O presente trabalho visa compreender o processo de degradação da parede celular vegetal de hospedeiros de fitopatógenos da família Xanthomonadaceae. Criamos e aperfeiçoamos uma técnica de enumeração dos genes relacionados ao metabolismo de polissacarídeos, com enfoque na distinção entre aqueles que agem sobre os componentes da parede celular vegetal e sobre outros polissacarídeos. A história evolutiva desse conjunto de enzimas foi delineada através de inferências sobre as relações de homologia entre os genes enumerados, sua presença ou ausência nos diversos genomas abordados e comparação das taxas de mutação entre grupos de homólogos. Além disso, procuramos também, com essa etapa de bioinformática e a etapa seguinte, incrementar a anotação desses genes, muitos descritos como hipotéticos ou com vaga definição de sua função. Na segunda parte do desenvolvimento do projeto foram feitos experimentos de expressão heteróloga e verificação da atividade enzimática para validação da anotação de alguns dos genes identificados. / This study aims to understand the process of degradation of host plant cell walls by plant pathogens of the Xanthomonadaceae family. We created and perfected a technique for enumeration of genes related to the metabolism of polysaccharides, focusing on the distinction between those who act on components of plant cell wall and on other polysaccharides. The evolutionary history of this group of enzymes has been outlined through inferences about the relations of homology between the genes listed, their presence or absence in different genomes and comparison of mutation rates between groups of homologues. Moreover, we also attempted with this bioinformatics step and the next step, to enhance the annotation of these genes, many described as hypothetical or vague in the determination of its function. In the second part of the project development heterologous expression and enzymatic activity assays were made to validate the annotation of some of the genes identified.

Retrovírus endógenos (ERVs) são vírus altamente difundidos no genoma de vertebrados. ERVs surgem quando retrovírus exógenos infectam células germinativas e se disseminam no genoma de seus hospedeiros, transmitindo seu material genético através das gerações por meio de herança mendeliana. ERVs são fundamentais na evolução dos genomas, sendo eles responsáveis por uma parte da diversidade genética de seus hospedeiros. O conhecimento sobre ERVs na família Felidae (Mammalia, Carnivora) estava praticamente restrito ao gato doméstico, e não se conhecia diversidade e padrões de evolução desses retroelementos em outras espécies. Este estudo teve como objetivo investigar diversidade, distribuição e padrões evolutivos de ERVs em espécies de gatos silvestres. Utilizando ferramentas de biologia molecular e bioinformática, foram identificadas e caracterizadas 85 sequências similares a retrovírus endógenos nos representantes das oito espécies brasileiras: Leopardus pardalis, L. wiedii, L. colocolo, L. geoffroyi, L. tigrinus, Puma concolor, P. yagouaroundi e Panthera onca. Encontrou-se uma predominância de ERVs similares a Gammaretrovirus, um padrão característico em muitas espécies de mamíferos. As análises filogenéticas evidenciaram três grupos principais de Gammaretrovirus, cada um evoluindo de maneira peculiar. Em uma visão geral, os ERVs provenientes de diferentes hospedeiros apresentaram-se distribuídos de forma heterogênea nas filogenias, dificultando a constatação de um padrão coevolutivo. No entanto, análises mais detalhadas de algumas sequências demonstraram peculiaridades, como no caso de um grupo de sequências similares a de um ERV oriundo do morcego Myotis lucifugus. Através de análises filogenéticas em comparação com dados obtidos na literatura, sugere-se que a infecção desse retrovírus ocorreu em uma espécie ancestral de felídeo, na segunda metade do Mioceno. Os resultados obtidos permitiram demonstrar que os felídeos neotropicais apresentam ERVs que seguem padrões semelhantes aos descritos a respeito de outros mamíferos, sugerindo também alguns casos de infecções de retrovírus muito similares entre diferentes ordens de mamíferos. / Endogenous retroviruses (ERVs) are widespread viruses in vertebrate genome. ERVs arise when exogenous retrovirus infects germinal cells and spread in the genome of their hosts, transmitting its genetic material throughout the generations by means of Mendelian inheritance. ERVs are fundamental for the evolution of genomes, being responsible for some part of the genetic diversity of their hosts. The knowledge on ERVs in felids (Mammalia, Carnivora, Felidae) was basically restricted to domestic cats, and the diversity and patterns of evolution of these retroviral elements in other species were not known. This study aimed to investigate diversity, distribution and evolutionary patterns of ERVs in wildcat species. Hence, by utilizing molecular biology and bioinformatics tools, 85 endogenous retrovirus-like sequences were identified and characterized in eight representative Brazilian species: Leopardus pardalis, L. wiedii, L. colocolo, L. geoffroyi, L. tigrinus, Puma concolor, P. yagouaroundi and Panthera onca. The analyses of these novel felid ERVs showed the predominance of Gammaretroviruslike sequences, which is a characteristic pattern present in many mammal species. Phylogenetic analyses have evidenced three major groups of Gammaretrovirus, each one evolving in a peculiar manner. ERVs from different hosts were distributed in a mixed way in the phylogenies, differently of a coevolutionary pattern. However, more detailed analyses of some sequences demonstrated peculiarities, as in the case of a group of sequences similar to an ERV from the bat Myotis lucifugus. Notably, through phylogenetic analyses, and in comparison to data obtained in the literature, it may be suggested that some infection by a retrovirus occurred in a felid ancestral species in the second half of the Miocene. Therefore, the results obtained demonstrate that ERVs from Neotropical felids follow patterns which are very similar to the ones described for other mammals, also suggesting some cases of similar retrovirus lineage infecting different mammal orders.

Os microRNAs (miRNAs) são pequenos RNAs não codificadores de proteínas presentes na maioria dos eucariotos. Esses RNAs regulam a expressão gênica em nível pós-transcricional através do silenciamento de mRNAs-alvo que possuem sítios complementares às suas sequências, atuando em praticamente todos os processos celulares. Embora a estrutura e função dos miRNAs estejam bem caracterizadas, aspectos relacionados à sua organização genômica, evolução e atuação em doenças são tópicos que apresentam enormes lacunas. Nesta tese, utilizamos abordagens computacionais para investigar estes temas em três trabalhos. No primeiro, processamos e integramos um vasto volume de dados publicamente disponíveis referentes aos miRNAs e genes codificadores de proteínas para cinco espécies de vertebrados. Com isso, construimos uma ferramenta web que permite a fácil inspeção da organização genômica dos miRNAs em regiões inter e intragênicas, o acesso a dados de expressão de miRNAs e de genes codificadores de proteínas (classificados em genes hospedeiros e não hospedeiros de miRNAs), além de outras informações pertinentes. Verificamos que a ferramenta tem sido amplamente utilizada pela comunidade científica e acreditamos que ela possa facilitar a geração de hipóteses associadas à regulação dos miRNAs, principalmente quando estão inseridos em genes hospedeiros. No segundo estudo, buscamos compreender como o contexto genômico e a origem evolutiva dos genes hospedeiros influenciam a expressão e evolução dos miRNAs humanos. Nossos achados mostraram que os miRNAs intragênicos surgem preferencialmente em genes antigos (origem anterior à divergência de vertebrados). Observamos que os miRNAs inseridos em genes antigos têm maior abrangência de expressão do que os inseridos em genes novos. Surpreendentemente, miRNAs jovens localizados em genes antigos são expressos em um maior número de tecidos do que os intergênicos de mesma idade, sugerindo uma vantagem adaptativa inicial que pode estar relacionada com o controle da expressão dos genes hospedeiros, e como consequência, expondo-os a contextos celulares e conjuntos de alvos diversos. Na evolução a longo prazo, vimos que genes antigos conferem maior restrição nos padrões de expressão (menor divergência de expressão) para miRNAs intragênicos, quando comparados aos intergênicos. Também mostramos possíveis associações funcionais relacionadas ao contexto genômico, tais como o enriquecimento da expressão de miRNAs intergênicos em testículo e dos intragênicos em tecidos neurais. Propomos que o contexto genômico e a idade dos genes hospedeiros são fatores-chave para a evolução e expressão dos miRNAs. Por fim, buscamos estabelecer associações entre a expressão diferencial de miRNAs e a quimioresistência em câncer colorretal utilizando linhagens celulares sensíveis e resistentes às drogas 5-Fluoruracil e Oxaliplatina. Dentre os miRNAs identificados, o miR-342 apresentou níveis elevados de expressão nas linhagens sensíveis à Oxaliplatina. Com base na análise dos alvos preditos, detectamos uma significativa associação de miR-342 com a apoptose. A superexpressão de miR-342 na linhagem resistente SW620 evidenciou alterações na expressão de genes da via apoptótica, notavelmente a diminuição da expressão do fator de crescimento PDGFB, um alvo predito possivelmente sujeito à regulação direta pelo miR-342. / MicroRNAs (miRNAs) are short non-coding RNAs found in most eukaryotic species. These RNAs regulate gene expression at post-transcriptional level by silencing target mRNAs through base-pairing of complementary sequences, thus acting on virtually all cellular processes. Although the structure and function of miRNAs are well understood, several aspects related to their genomic organization, evolution and involvement with diseases are largely underexplored. In this thesis, we employed computational methods to investigate such issues in three different studies. In the first one, we have processed and integrated a large amount of public data related to miRNAs and coding genes for five vertebrate species. Then, we developed a webtool to allow the analysis of the miRNA genomic context in inter and intragenic regions, the access of miRNA and gene expression data (classified as host and non-host genes), as well as other relevant information. We noticed that the webtool has been largely used by the scientific community, and we believe that it can facilitate hypothesis generation related to miRNA regulation, especially when they are within host genes. In the following study, we sought to understand how the genomic context and the evolutionary origin of host genes can affect the expression and evolution of human miRNAs. Our findings showed that intragenic miRNAs are preferentially embedded within old host genes (originated before the split of vertebrates). We observed that miRNAs within old genes are more broadly expressed than those within young genes. Surprisingly, young miRNAs within old genes were expressed in more tissues than their intergenic counterparts, suggesting an initial adaptive advantage which might be related to their hosts expression control, and as a consequence, exposing them to a more diverse cellular contexts and target genes. In the long run, we found that old host genes lead to expression constraints (lower expression divergence) between species for intragenic miRNAs, in respect to intergenic ones. We also showed possible functional associations related to miRNA genomic context, such as the enrichment of young intergenic miRNAs in testis, while young intragenic miRNAs were enriched in neural tissues. Thus, we propose that the genomic context and the age of the host genes are key factors in shaping the expression and evolution of miRNAs. Finally, we sought to establish associations between differential expression of miRNAs and chemoresistance in colorectal cancer using resistant and sensitive cell lines to 5-Fluoruracil and Oxaliplatin. Among differentially expressed miRNAs, miR-342 was highly expressed in sensitive cell lines to Oxaliplatin. Based on target prediction analysis, miR-342 is likely associated with apoptosis. The induced overexpression of miR-342 in SW620, a cell line resistant to Oxaliplatin, changed the expression levels of genes linked to the apoptosis pathway, notably the downregulation of PDGFB growth factor, which is a predicted target possibly subjected to direct regulation by miR-342.

Les récents progrès des technologies de séquençage et des techniques de biologie moléculaire ont permis aux chercheurs de révéler l'ubiquité et la diversité des virus. Malgré ces progrès, la découverte des virus s'est surtout concentrée sur les virus de vertébrés. De plus, les virus connus sont très peu étudiés en terme d'écologie virale. Les arthropodes de par leur histoire évolutive ancienne représentent 80% de la diversité animale sur notre planète. L'omniprésence de l'endosymbiose chez les arthropodes et leur capacité d’héberger des symbiotes (parfois multiples) suggèrent que leurs interactions avec leurs virus pourraient également être de nature symbiotique, allant des interactions durables de type mutualiste à celles de type antagoniste. Dans ce contexte, mon travail de thèse s’est concentré sur un densovirus de moustiques, le Culex pipiens densovirus (CpDV), en étudiant leur prévalence et leur diversité dans les populations naturelles de moustiques Culex pipiens et leurs interactions avec les bactéries endosymbiotiques Wolbachia dans un système naturel hôte-bactérie-virus (Cx. pipiens -Wolbachia - CpDV), tant dans des populations naturelles que des colonies de laboratoire. Nous avons révélé une forte prévalence et une grande diversité de CpDV dans les populations mondiales de Cx. pipiens. De plus, le CpDV infecte de façon persistante les lignées de laboratoire et est transmis verticalement avec l'endosymbiote Wolbachia. Nous montrons une prévalence et une diversité élevées du CpDV en Turquie et en Tunisie, et nous nous sommes concentrés davantage sur ces deux pays pour étu-dier les interactions Wolbachia-CpDV dans les populations naturelles. En Tunisie, dans une zone de contact étroite où deux groupes de Wolbachia génétiquement distincts coexistent, ces différents groupes de Wolba-chia influencent la prévalence et la diversité du CpDV. Nous montrons également une corrélation positive entre la densité de Wolbachia et la densité de CpDV. D’une manière globale, nos résultats suggèrent que le système CpDV-Cx.pipiens -Wolbachia et plus généralement les densovirus sont de bons modèles pour étudier les interactions écologiques entre ces partenaires et leur évolution dans la nature et au laboratoire. / Virus discovery has long depended on the observation of outbreaks, and the research to understand their pathogenicity depended on their culture. The advances of sequencing technology and molecular biology tech-niques allowed researchers to discover the ubiquity and the diversity of viruses. Despite these advances, virus discovery remains mainly focused on vertebrate viruses. Furthermore, even the known diversity of these virus-es is poorly studied in term of virus ecology. Arthropods with their ancient evolutionary history represent 80% of animal diversity on our planet. The ubiquity of endosymbiosis in arthropods and their ability to support (sometimes multiple) symbionts suggest that their interactions with their viruses might be of symbi-otic nature, intimate long-term associations with outcomes ranging from mutualistic to antagonistic. In this context, this PhD dissertation focuses on a mosquito densovirus, Culex pipiens densovirus (CpDV), to in-vestigate their prevalence and diversity in natural populations of Culex pipiens mosquitoes and their inter-actions with endosymbiotic bacteria Wolbachia in a natural host-bacteria-virus system (Cx. pipiens -Wolbachia - CpDV) both in natural populations and laboratory colonies. We reveal a high prevalence and diversity of CpDV in worldwide Cx. pipiens populations. Moreover, CpDV persistently infects laboratory lines and are vertically transmitted along with their hosts’ endosymbiont Wolbachia. We reveal high CpDV prevalence and diversity in Turkey and Tunisia and further focus on these two countries to investigate Wolbachia-CpDV interactions in natural populations. In Tunisia, in a narrow contact zone where two ge-netically distinct Wolbachia groups coexist, different Wolbachia groups influences CpDV prevalence and diversity. Moreover, we show a positive correlation between Wolbachia density and CpDV density. Overall, our results suggests that CpDV-Cx.pipiens -Wolbachia system is a good model system to study the ecologi-cal interactions between these partners and their evolution together both in the nature and in laboratory conditions. Densoviruses with their diversity and long-term evolution with their hosts, and their easily ma-nipulated small genomes, are good models to study complex host-virus-bacteria interactions as well as virus ecology and evolution in a symbiotic context.

A família gênica Forkhead P {FOXP) tem sido alvo de muitos estudos envolvendo evolução do cérebro e comportamento animal. Destacam-se particularmente as investigações com o gene FOXP2, que indicam que mudanças neste gene estariam associadas com a evolução da vocalização em algumas espécies de mamíferos, incluindo o Homo sapiens. Recentemente, estudos de desordem intrínseca de proteínas (IDPs) tem ganhado ênfase no contexto evolut ivo, visto que uma correlação posit iva entre regiões de desordem e altas taxas evolutivas tem sido observada. Através de um conjunto de abordagens que inclui predizer o conteúdo de desordem e os motivos lineares de interação, bem como as taxas evolutivas, buscamos desvendar a historia evolutiva dos genes da subfamília FOXP. Concentramos nossas análises sobre regiões desordenadas das proteínas FOXPl, FOXP2, FOXP3 e FOXP4 encontradas em 77 espécies de tetrápodes. Tais regiões proteicas são normalmente negligenciadas em estudos dessa natureza, pois se localizam fora de seus tra dicionais domínios conservados, normalmente associados à função principal da proteína. Sít ios apontados estando sob seleção positiva e relaxamento da restrição seletiva mostraram-se hotspots importantes para mudanças que podem impactar na capacidade de interação das proteínas. Encontramos que os maiores valores de w são mais prevalentes em regiões desordenadas que em ordenadas. Ainda, alto e similar valor de desordem (70%) foi encontrado nas 77 proteínas ortólogas de FOXPl , FOXP2, e FOXP4, indicando a manutenção de um "padrão geral" sobre um longo tempo evolutivo. Portanto, a variabilidade tanto de aminoácidos quanto de motivos lineares dentro das regiões de desordem foi marcante. A proteína FOXP3 apresentou menor nível de desordem (30%), mas signif icante sinal de seleção positiva em alguns sítios. Composição idênt ica de resíduo de aminoácido e/ou motivos lineares em espécies filogeneticamente distantes, indica clara convergência molecular, provavelmente associada a pressões seletivas similares. Sucessivamente, nossos achados mostraram uma clara diferença na composição de motivos lineares entre mamíferos e não mamíferos, dando suporte para a importância dos estudos de evolução da interatividade proteica para as compreensões de características taxa-específicas. / Forkhead Family P (FOXP) has been target of many studies about brain and behavior evo lution among species. FOXP2 receives special attention in academic society, due associations with vocalízation evolution in mammals, including Homo sapiens. Recently, intrinsically disorder proteins studies have gained emphasis in the evolutionary context, as positive correlation between disorder regions and higher evolutionary rate has been observed. Through a set of approaches, including disorder and linear motif predictions, as well as estimate evolutionary rates, we aimed to unveil the evolutionary history of FOXP subfamily genes. We focused our ana lysis over disordered regions of FOXPl, FOXP2, FOXP3 and FOXP4 proteins retrieved in 77 tetrapods. Such protein regions are usually neglected in studies of this nature, for being localized out of the traditional conserved domains, usua lly associated with the main function of the protein. Sites indicated as under relaxation of selective constrains or positive selection have shown to be important hotspots for changes that can impact in protein interaction capability. Higher w va lues are prevalent in disordered regions than in ordered ones. Still, high and similar disorder proportion (~70%) was found among 77 orthologues proteins of FOXPl, FOXP2 and FOXP4, indicating general pattern of disorder maintenance, along tetrapod's evolutionary tree. However, amino acid and linear motifs variability within disordered regions was observed. FOXP3 protein presented lower disorder leveis (~30%), when compared with other paralogues, but signal of positive selection was observed in some sites. ldentical composition of amino acid residues and/or linear motifs is, probably, associated with similar selective pressure. Successively, ou r results showed clear differences in linear motif composition between mammals and non-mammals, supporting the importance of evolutionary studies on protein interaction for the understanding of taxa-specifics characteristics.

Submitted by Sandro Camargo (sandro.camargo@unipampa.edu.br) on 2015-03-08T19:40:44Z No. of bitstreams: 1 117110035.pdf: 2502976 bytes, checksum: 4587ac7502f70421540e27b087febdfb (MD5) / Made available in DSpace on 2015-03-08T19:40:44Z (GMT). No. of bitstreams: 1 117110035.pdf: 2502976 bytes, checksum: 4587ac7502f70421540e27b087febdfb (MD5) Previous issue date: 2013-08-05 / A oxitocina e a vasopressina são nonapeptídeos intimamente relacionados que surgiram a partir de duplicações em tandem ocorridas no ancestral comum dos vertebrados mandibulados e que desempenham importantes funções fisiológicas e comportamentais em muitos organismos. Suas atividades são mediadas através das interações com o receptor da oxitocina (OXTR) e com os receptores da vasopressina (AVPR1a, AVPR1b e AVPR2). O objetivo do presente estudo é caracterizar a evolução molecular dessa família de receptores num conjunto grande de organismos. Assim como, avaliar a variabilidade genética de OXTR na espécie Ovis aries que ainda não possui seu genoma disponível nos bancos de dados. Devido à conservação dos nonapeptídeos, ao menos considerando os mamíferos, acredita-se que a evolução do sistema esteja ocorrendo através de seus receptores, que, portanto são o centro deste estudo. Através de sequências nucleotídicas e proteicas para todos os receptores da oxitocina e da vasopressina obtidas nos bancos de dados HMMER e Uniprot foram realizadas análises filogenéticas por máxima verossimilhança no programa Mega 5.0. Os dados disponíveis no site Genomicus foram utilizados para a realização da sintenia e vizinhança. O padrão de evolução molecular foi estabelecido utilizando um conjunto de 23 espécies de mamíferos usando o pacote Paml 4.6. A fim de contribuir com o conhecimento da espécie Ovis aries analisou-se o gene OXTR, na raça Crioula Lanada (N=35) que apresenta um padrão distinto de seleção em comparação com as raças Ile de France (N=6) e Ideal (N=5) que são selecionadas para corte e lã. Nossos resultados indicam que OXTR, AVPR1a, AVPR1b, AVPR2 e o bloco sintênico desses receptores emergiram com o ancestral comum dos vertebrados no primeiro evento de duplicação do genoma. Primeiramente foram originados, o gene ancestral de AVPR1a e AVPR1b e o gene ancestral de OXTR e AVPR2. Os quatro receptores observados hoje se originaram após o segundo evento de duplicação do genoma ter ocorrido no ancestral comum dos vertebrados. Em termos de evolução molecular foram encontrados padrões distintos entre animais domesticados e selvagens nos receptores AVPR1a e AVPR1b. Isso indica que os animais domesticados estão sob seleção positiva para esses genes, clara marca molecular que pode estar ligada ao processo de seleção para docilidade, num contexto de evolução convergente. Para a espécie Ovis aries foi amplificado um total de 1644 pares de base (pb) obtidos do DNA das três raçasovinas estudadas. Embora tenha sido registrada variação entre as raças, não há uma quebra no padrão de evolução neutro com sinal de seleção purificadora. O que reforça a ideia que o sistema todo esteja evoluindo via os dois receptores de AVP, cuja origem filogenética é comum, AVPR1a e AVPR1b. / Oxytocin and vasopressin are closely related nonapeptides that arose from tandem duplication occurred in the common ancestor of jawed vertebrates and play important physiological and behavioral functions in many organisms. Its activities are mediated through interactions with the oxytocin receptor (OXTR) and vasopressin receptors (AVPR1a, AVPR1b and AVPR2). The aim of this study is to characterize the molecular evolution of this family of receptors in a large set of organisms. As well as, assess the genetic variability of the OXTR from Ovis aries species that does not have its genome available in databases. Due to the conservation of nonapeptides, at least considering the mammals, it is believed that the evolution of the system is going through their receptors, which therefore are the focus of this study. Through nucleotide and protein sequences for all oxytocin and vasopressin receptors obtained in databases UniProt and HMMER, phylogenetic analyzes were performed by maximum likelihood in the Mega 5.0 program. Available data on the website Genomicus were used to carry out the synteny and neighborhood. The pattern of molecular evolution was established using a set of 23 mammalian species using the Paml 4.6 package. To contribute to the knowledge of the Ovis aries species it was analyzed the OXTR gene, in the Creole Lanada breed (N = 35) that presents a distinct pattern of selection compared to the breeds Ile de France (N = 6) and Ideal (N = 5) that are selected for cutting and wool. Our results indicate that OXTR, AVPR1A, AVPR1b, AVPR2 and the sintenic block of these receptors emerged with the common ancestor of vertebrates in the first genome duplication event. First, was originated the ancestral gene of AVPR1A and AVPR1b and the ancestral gene of OXTR and AVPR2. The four receptors observed today originated after the second genome duplication event occurred in the common ancestor of vertebrates. In terms of molecular evolution were found distinct patterns between domesticated and wild animals in the AVPR1A and AVPR1b receptors. This indicates that domesticated animals are under positive selection for these genes, clearly molecular mark which may be linked to the process of selection for docility, in the context of convergent evolution. For the Ovis aries species was amplified a total of 1644 base pairs (bp) of DNA obtained from three sheep breeds studied. Although has been found variation between the breeds, there is no break in the pattern of neutrality with signal of purifying selection. This strengthens the idea that the whole system is evolvingthrough the two AVP receptors, whose phylogenetic origin is common, AVPR1A and VPR1b.

In this study, I sequenced the mitochondrial cytochrome b gene to elucidate the extent and pattern of genetic variations among and within populations of Rhinichthys osculus (Santa Ana Speckled Dace) found in the different watersheds in Southern California, Central California Coast and Eastern California Desert. I described and analyzed the structural characteristics and pattern of base sequence substitutions in the cytochrome b gene to understand the molecular evolution of the gene. The Phylogenetic analyses showed that the Southern California Santa Ana Speckled dace is a distinct population from the Central California Coast dace population and Eastern California Desert dace population, and is more closely related to the Colorado River speckled dace population. There is a high degree of genetic variation among all populations including a significant genetic structure associated with watersheds, mountain ranges, and geographic grouping based on locations. The SWISS-Model automated protein structure homology elucidated the conserved and invariant residues within the cytochrome b gene where the amino acid substitutions are located in the trans-membrane of the protein sequence. The implication for conservation and management of the Southern California Santa Ana Speckled dace is high because of habitat lost for this distinct dace population. The data from this study will contribute to preserving the genetic variability of the Santa Ana Speckled Dace as a separate taxa and species, as well as to help maintain intact the population in the different Southern California creeks.

The trajectory of evolutionary adaptation can be influenced both by the interactions of organisms with their environments as well as by the biological characteristics of the organisms themselves. My dissertation research uses the New Zealand freshwater snail Potamopyrgus antipodarum to 1) gain important insight into how coevolutionary interactions between hosts and parasites influence patterns of gene expression and genetic differentiation of hosts and, 2) evaluate how reproductive mode, and ploidy level affect patterns of adaptive molecular evolution. Coevolutionary interactions between hosts and parasites are a primary source of strong natural selection that can lead to rapid evolutionary change. Here, I used evaluation of patterns of gene expression and genetic differentiation to take critical steps towards characterizing the genomic basis of coevolutionary interactions between P. antipodarum and Microphallus livelyi. I found that M. livelyi-infected P. antipodarum exhibit systematic downregulation of genes relative to uninfected P. antipodarum. The specific genes involved in response to parasites differ markedly across lakes, consistent with population-specific host-parasite interactions leading to population-specific evolutionary trajectories. I also identified a set of rapidly evolving loci that represent promising candidates for targets of parasite-mediated selection across lakes as well as within each lake population. These results constitute the first genomic evidence for population-specific responses to coevolving infection in the P. antipodarum-M. livelyi interaction and provide new insights into the genomic basis of coevolutionary interactions in nature. I also generated and characterized the first transcriptomic resources for Microphallus parasites collected from two species of Potamopyrgus snails (P. antipodarum and P. estuarinus). These data both revealed that these parasites appear to represent distinct genetic lineages, which is interesting in light of the tight coevolutionary interactions between P. antipodarum and M. livelyi, and lay the groundwork for future research. Polyploidy has the potential to facilitate adaptive evolution by providing redundant genome copies that are free to evolve new functions. By contrast, asexuality, with which polyploidy is often associated, is expected to restrict adaptive evolution by decreasing the efficacy of natural selection and access to new genetic variation. I evaluated whether and how ploidy level and reproductive mode influence patterns of adaptive molecular evolution in P. antipodarum to assess 1) the potential evolutionary genomic benefits of recent polyploidy, and 2) how patterns of adaptive molecular evolution in asexuals are influenced by polyploidy. I compared patterns of positive selection in 60 genes across 27 P. antipodarum lineages (10 diploid sexuals, 12 triploid asexuals, 5 tetraploid asexuals) and a diploid sexual outgroup, Potamopyrgus estuarinus. I found little evidence that ploidy level and/or reproductive mode influence patterns of positive selection in P. antipodarum. Even so, this study provides initial steps in evaluating whether ploidy level and reproductive mode influence patterns of adaptive molecular evolution. Taken together, my dissertation work contributes new insights to the field of host-parasite coevolutionary interactions and will inform future studies into how ploidy level and reproductive mode influence patterns of adaptive molecular evolution.