• Refine Query
  • Source
  • Publication year
  • to
  • Language
  • 69
  • 6
  • 6
  • 6
  • 6
  • 6
  • 6
  • 4
  • 3
  • 3
  • 3
  • 2
  • 1
  • 1
  • Tagged with
  • 116
  • 116
  • 116
  • 30
  • 24
  • 22
  • 17
  • 17
  • 17
  • 16
  • 13
  • 12
  • 12
  • 12
  • 12
  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
71

Mechanisms of translation regulation in long-term synaptic plasticity

Hebert-Seropian, Sarah 12 1900 (has links)
Les souvenirs sont encodés dans le cerveau grâce aux configurations uniques de vastes réseaux neuronaux. Chaque connexion dans ces circuits est apte à être modifiée. Ces changements durables s’opèrent au niveau des synapses grâce à une synthèse de protéines de novo et génèrent ce qu’on nomme des traces mnésiques. Plusieurs preuves indiquent que, dans certaines formes de plasticité synaptique à long terme, cette synthèse a lieu dans les dendrites près des synapses activées plutôt que dans le corps cellulaire. Cependant, les mécanismes qui régulent cette traduction de protéines demeurent encore nébuleux. La phase d’initiation de la traduction est une étape limitante et hautement régulée qui, selon plusieurs chercheurs, constitue la cible principale des mécanismes de régulation de la traduction dans la plasticité synaptique à long terme. Le présent projet de recherche infirme cette hypothèse dans une certaine forme de plasticité synaptique, la dépression à long terme dépendante des récepteurs métabotropiques du glutamate (mGluR-LTD). À l’aide d’enregistrements électrophysiologiques de neurones hippocampiques en culture couplés à des inhibiteurs pharmacologiques, nous montrons que la régulation de la traduction implique les étapes de l’élongation et de la terminaison et non celle de l’initiation. De plus, nous démontrons grâce à des stratégies de knockdown d’expression d’ARN que la protéine de liaison d’ARNm Staufen 2 joue un rôle déterminant dans la mGluR-LTD induite en cultures. Dans leur ensemble, les résultats de la présente étude viennent appuyer un modèle de régulation de la traduction locale de protéines qui est indépendante de l’initiation. / Memories are encoded in the unique configurations of the vast neuronal networks of the brain. Each of these connections possesses the ability to be modified. Such long-lasting changes at the synapse often require the synthesis of new proteins that create what we call memory traces. Evidence suggests that the signal-induced activation of translation in some forms of synaptic plasticity occurs locally, at the activated synapses, rather than in the soma. However, the mechanisms regulating local and rapid de novo protein synthesis are poorly understood. The initiation step of translation is a highly regulated step and is believed to be the main target of control. The present research project challenges this view for a certain form of long-term synaptic plasticity, metabotropic glutamate receptor-dependent long-term depression (mGluR-LTD). We show, using electrophysiological recordings of dissociated hippocampal neurons in cultures coupled to pharmacological inhibitors, that translation regulation depends on elongation and termination, rather than initiation. Moreover, by exploiting RNA knockdown strategies, we demonstrate that the RNA-binding protein Staufen 2 plays a crucial role in mGluR-LTD induced in cultures. Altogether, the findings of the present study support a model of translation regulation that is downstream of initiation.
72

Functional characterization of the FET family of RNA-binding proteins

Baethge, Kerstin 03 July 2014 (has links)
RNA-bindende Proteine spielen eine zentrale Rolle in der posttranskriptionellen Kontrolle von mRNAs, die zwischen Transkription und Abbau von mRNAs stattfindet. RNA-bindende Proteine beeinflussen Spleißen, Export, Stabilität, Lokalisierung und Translation von mRNAs. FUS, EWSR1 und TAF15 gehören zu der Familie der FET Proteine. Diese wirken an verschiedenen zellulären Prozessen wie Transkription, Spleißen und der Prozessierung von miRNAs mit. Translokationen und Mutationen der FET Proteine führen zu verschiedenen Krankheiten. FUS spielt eine Rolle bei den neurodegenerativen Krankheiten frontotemporale Lobärdegeneration (FTLD) und amyotrophe Lateralsklerose (ALS). In dieser Arbeit wurde die mithilfe von photoaktivierbaren Ribonukleotiden UV-Licht induzierte Quervernetzung und Immunpräzipitation (PAR-CLIP) Methode genutzt, um die RNA-Bindestellen von FUS, EWSR1 und TAF15, einer ALS-verursachenden FUS Mutante und einem anderen, mit ALS in Verbindung stehenden Protein, TARDBP, zu bestimmen. Die RNA-Bindestellen der FET-Proteine lagen größtenteils in Introns. Passend dazu konnte durch knockdown der FET Proteine eine Rolle von FUS und EWSR1 im Spleißen von mRNAs validiert werden. Dem Ubiquitin-Proteasom-System zugehörige RNAs waren unter den sowohl von FUS als auch TARDBP gebundenen mRNAs überrepräsentiert. Dies bestätigt die Annahme, dass Störungen in der Proteindegradation die ALS-Pathogenese beeinflussen. Zusätzlich konnte gezeigt werden, dass FUS und TAF15 bevorzugt UAC-reiche, einzelsträngige RNA-Sequenzen binden. Sequenzierung von mRNAs nach Depletion von FUS, EWSR1 und TAF15 in HEK293-Zellen zeigte einen stabilisierenden Effekt der FET-Proteine auf gebundene mRNAs. Desweiteren scheinen die FET Proteine durch Interaktion mit Promotor-assoziierten, nicht-kodierenden RNAs die Transkription zu beeinflussen. / Post-transcriptional regulation of gene expression takes place at multiple levels between transcription and decay of the mRNA. RNA-binding proteins play a key role in orchestrating splicing, export, stability, localization and translation of mRNAs. FUS, EWSR1 and TAF15 constitute the FET protein family which participates in multiple levels of cellular function. FET proteins have been implicated to function in various cellular processes including transcription, pre-mRNA splicing and miRNA processing. Translocations and mutations in FET proteins lead to diverse pathologies. FUS is involved in neurodegenerative diseases like frontotemporal lobar degeneration (FTLD) and amyotrophic lateral sclerosis (ALS). In this study, Photoactivatable-Ribonucleoside-Enhanced Crosslinking and Immunoprecipitation (PAR-CLIP) was used to determine RNA-targets and binding sites of FUS, EWSR1 and TAF15, an ALS-causing FUS mutant and another ALS-related protein, TARDBP. The identified binding sites of FET proteins were mainly intronic, supporting the involvement of FUS and EWSR1 in splicing, which was validated by FET protein knockdown. Comparison of FUS and TARDBP RNA targets revealed that ubiquitin-proteasome related gene categories were overrepresented, further illustrating that aberrations in protein degradation are implicated in the pathogenesis of ALS. In addition, it was shown that FUS and TAF15 proteins preferentially bind UAC rich, single-stranded RNA sequences. mRNA sequencing after FUS, EWSR1 and TAF15 depletion in HEK293 cells revealed a stabilizing effect on their targets. Interestingly, FET proteins also seem to influence transcription by interaction with promoter-associated noncoding RNAs. In summary, we identified the RNA-targets and binding sites of all human FET proteins in comparison with an ALS-causing FUS mutant and TARDBP. Functional studies revealed an involvement of FET proteins in mRNA stabilization, splicing and transcriptional regulation.
73

RNA-Bindestudien und funktionelle Analysen der chloroplastidären RNA-Bindeproteine CP29A und CP31A in Arabidopsis thaliana

Kupsch, Christiane 09 January 2014 (has links)
cpRNPs (chloroplastidäre Ribonukleoproteine) sind kernkodierte RNA-Bindeproteine, die jeweils zwei RRM (RNA recognition motif)-Domänen besitzen und in die Chloroplasten höherer Landpflanzen importiert werden. Die Mitwirkung dieser Proteine an der Reifung plastidärer mRNAs in vitro, ihre bemerkenswert hohe Abundanz sowie die Beeinflussung ihrer Funktion durch lichtabhängige posttranslationale Modifikationen weisen auf eine bedeutende Rolle der cpRNPs in der Regulierung der plastidären Genexpression hin. In dieser Arbeit erfolgte erstmals eine Analyse der in vivo bestehenden Interaktionen zwischen cpRNPs und RNA im Stroma von Chloroplasten mittels RIP-Chip-Analysen. Es konnte gezeigt werden, dass ein Großteil der plastidären mRNA-Spezies mit CP31A und CP29A kopräzipitiert. Neben der mehrheitlichen Überlappung der RNA-Interaktionen konnten spezifische Präferenzen von CP29A und CP31A für bestimmte Transkripte identifiziert werden. Um Einblicke in die molekularen Funktionen der cpRNPs zu erhalten, wurden cp29a- und cp31a- Einzel- und Doppelmutanten hinsichtlich der Akkumulation, sowie des Spleiß- und Edierungsstatus plastidärer mRNAs untersucht. Da unter Standard-Wachstumsbedingungen lediglich wenige milde Defekte innerhalb der mRNA-Reifung in den Mutanten detektiert wurden, erfolgten weitere Analysen unter Kältestress-Bedingungen, die zu einer Induktion der CP29A-Expression und zu dem Ausbleichen junger Blattgewebe in cp29a- und cp31a-Mutanten führten. In diesen chlorotischen Geweben wurden reduzierte Akkumulationen plastidärer Proteine sowie PEP-abhängig transkribierter mRNAs detektiert. Darüber hinaus Prozessierungsdefekte für einige Transkripte beobachtet: Die Reifung der 23S-rRNA findet in cp29a- und cp31a-Mutanten nur eingeschränkt statt. In cp31a-Mutanten sind zudem die Prozessierung der ycf3-RNA sowie die Spaltung eines polycistronischen rpl33-Vorläufers gestört. In cp29a-Mutanten ist unter anderem die Prozessierung der rpoC1- und der ycf3-mRNA defizitär. / cpRNPs (chloroplast ribonucleoproteins) are nuclear encoded RNA-binding proteins, which contain two RRM (RNA recognition motif) domains and reside within the chloroplasts of higher land plant species. They are involved in plastid mRNA accumulation and processing in vitro. Their involvement in multiple steps of mRNA maturation and their remarkable abundance together with their regulation via posttranslational modifications identified the cpRNPs as potential central regulators of chloroplast gene expression. This work investigated the in vivo interactions of CP29A and CP31A from Arabidopsis with RNA in isolated chloroplasts via RIP-Chip. In this approach the association of both proteins with the majority of plastid mRNA species could be shown. While the interaction profiles of CP29A and CP31A were largely overlapping, also specific preferences for some transcripts were detected. To gain insights in the molecular functions of CP29A and CP31A, null mutants for one or both cpRNPs were analyzed for mRNA accumulation, splicing and editing. Since only few mild mRNA maturation defects were detected at standard growth conditions, analyses at cold stress were performed. Cold stress leads to an increase in CP29A expression and in both cp29a- and cp31a single- and double mutants a bleaching of young leaf tissue was observed. The chlorotic tissues showed a strong decrease in plastid protein accumulation accompanied by reduced levels of PEP-dependent transcripts. In addition some processing defects were observed: The maturation of the 23S-rRNA was reduced in cp29a- and cp31a mutants. In addition the processing of the ycf3-mRNA and a polycistronic rpl33 transcript were defective in cp31a mutants. In cp29a mutants processing defects within the rpoC1- and the ycf3-mRNA were detected.
74

Funktion und Evolution chloroplastidärer PPR-Proteine

Beick, Susanne 16 May 2011 (has links)
PPR-Proteine bilden die größte Familie von RNA-Bindeproteinen in Pflanzen und sie werden fast ausschließlich in die Mitochondrien oder Plastiden importiert, wo sie eine wesentliche Rolle im RNA-Metabolismus spielen (Lurin et al., 2004). Doch die Funktionsweise der Proteine ist noch weitgehend unbekannt. In dieser Arbeit wurde das plastidäre PPR-Protein PPR5 in Zea mays funktionell charakterisiert, dessen Ortholog in Arabidopsis thaliana essentiell für die Embryogenese ist (Cushing et al., 2005). Mittels PPR5-Immunopräzipitation und einer Analyse der kopräzipitierten RNA konnte in vivo eine spezifische Assoziation mit der ungespleißten tRNA-Glycin (UCC) nachgewiesen werden. Analysen von ppr5-Mais-Mutanten offenbarten einen Stabilitätsverlust dieser RNA. Es wurde gefolgert, dass PPR5 das Transkript vor einem endonukleolytischen Abbau schützt. Die weiteren Projekte der Arbeit widmeten sich der Evolution der Familie. Um Erkenntnisse zur Funktion und Spezifität nahe verwandter PPR-Proteine zu erhalten, wurden die drei nächsten Verwandten von PPR5 identifiziert und Mais-Mutanten isoliert. Weiterhin wurde PPR54 untersucht. Es konnte gezeigt werden, dass PPR54 in Mais – wie in Arabidopsis (Tillich, nicht publiziert) – für das Spleißen des ndhA-Introns benötigt wird. Damit wurden erstmalig orthologe PPR-Proteine in einer Mono- und einer Dikotylen funktionell analysiert. Die vorgelegten Analysen mündeten in drei allgemeingültigen Schlussfolgerungen zur Funktion der PPR-Proteine. 1) Plastidäre PPR-Proteine, die in Dikotylen wie Arabidopsis für die Embryogenese notwendig sind, üben eine Funktion in der plastidären Translation aus. 2) Die vorgeschlagene Funktionsweise von PPR5 erfordert nicht die Rekrutierung anderer, katalytisch aktiver Proteine, sondern ihr liegt ein passiver, auf der Bindung einer RNA beruhender Mechanismus zugrunde. 3) Die Funktion orthologer PPR-Proteine ist in Mono- und Dikotylen konserviert, wie am Beispiel von PPR54 experimentell nachgewiesen wurde. / PPR proteins are the largest family of RNA binding proteins in plants and the vast majority of them is localized to mitochondria or chloroplasts, where they are major players in the RNA metabolism of defined transcripts (Lurin et al., 2004). However, the mechanistic function of these proteins is still not clear. In this study, the plastid PPR protein PPR5, whose ortholog in Arabidopsis thaliana is embryo-essential (Cushing et al., 2005), was functionally characterized in Zea mays. By PPR5 immunoprecipitation and analyses of the coimmunoprecipitated RNA, a specific association to the unspliced tRNA glycine (UCC) was shown in vivo. The analysis of ppr5 maize mutants demonstrated a loss of stability of the tRNA precursor in mutants. It was concluded that the interaction with PPR5 protects the unspliced tRNA from endonucleolytic decay. In addition, close relatives of PPR5 were identified in maize (PPR2, PPR50, and PPR51) by phylogenetic means and maize mutants were isolated. A future characterization of four paralogous PPR proteins might answer whether closely related PPR proteins have similar functions or RNA targets. The analysis of PPR54 in maize demonstrated that PPR5 is needed for the splicing of the ndhA intron in maize as it is in Arabidopsis (Tillich, not published). Three important conclusions concerning the function of PPR proteins in general were drawn from the studies of chosen PPR proteins presented here. First, plastid PPR proteins that are essential in embryo development in eudicots like Arabidopsis should be necessary for plastid translation in most cases. Second, the characterization of PPR5 revealed a possibly ancient functional mechanism of PPR proteins which does not invoke the recruitment of additional catalytic factors but relies on the passive binding of RNA elements. Last, the conservation of function of orthologous PPR proteins in monocots and eudicots, which was shown in the case of PPR54, was demonstrated experimentally for the first time.
75

Functional characterization of a Krüppel zinc finger protein- zinc finger protein 146. / CUHK electronic theses & dissertations collection

January 2008 (has links)
By means of reverse-transcription polymerase chain reaction, overexpression of ZNF146 was detected in two human HCC cell lines HepG2 and Hep3B and a clear relationship between HCC and overexpression of ZNF146 has been established. Subcellular localization of ZNF146 protein in liver cells was studied by generation and expression of a green fluorescent protein (GFP) fusion protein. The nuclear localization and the reported DNA binding ability of ZNF146 protein provided a hint that ZNF146 may carry out its function in the cell system by interacting with specific genomic DNA sequences. Recombinant ZNF146 protein was expressed using bacterial and yeast system for the genomic DNA pull down assay in the identification of potential interacting genomic DNA sequences. Several potential genomic DNA sequences that interact with ZNF146 were identified and the gene MDM2 is the one of the candidates that is directly related to human carcinogenesis. MDM2 is a negative regulator of the tumor suppresser protein p53. Deregulation of MDM2 will impair the cell's ability in cell cycle arrest, DNA repair and apoptosis upon induced DNA damage. / Hepatocellular carcinoma (HCC) is a type of primary malignant liver tumor. And is one of the most frequent malignancies worldwide. The focus of this research project is the characterization of a Kruppel zinc finger protein, zinc Finger Protein 146 (ZNF146) using HCC as a disease model. The aim of this project is to understand the functional role ZNF146 and try to explore the mechanism of how ZNF146 might be involved in the carcinogenesis of HCC. / In order to have a better understanding with the protein ZNF146, SUMOylation properties of this protein has been studied. SUMO1 modification on ZNF146 has already been reported. And in our study, experimental result demonstrated that ZNF146 is also modified by SUMO2 and SUMO3 in liver cells. Other than the SUMOylation sites for SUMO1 protein which has been reported, modification sites for SUMO2 at the K247 and K275 positions were mapped, while K191R, K219R, K247R, K256R and K275R, five positions were mapped for SUMO3 modification. A more complete picture of the SUMOylation properties of ZNF146 has been revealed. Since we hypothesized that ZNF146 is related to the p53 tumor suppressor, cell cycle control and DNA repair pathway, a cell cycle study using flow cytometry was performed for the investigation of the effect on cell cycle regulation by ZNF146 overexpression. In our study, ZNF146 overexpression promoted the G1/S transition in the cell division cycle, which indicated that liver cells were more active for the progression of cell cycle. / On the other hand, using cDNA microarray technology expression profiles of ZNF146 overexpressing and non-overexpressing liver cell lines were compared and with real-time polymerase chain reaction, six candidate genes CRLF1, IFI44, ST6GAL1, LOC441601, IL18 and RAD17 were confirmed with their deregulation induced by the overexpression of ZNF146. Four of the candidates, IFI44, LOC441601, IL18 and RAD17 were found to be related to the p53 tumor suppressor activity or DNA damage, repair response and control. This observation, together with the result of genomic DNA pull down assay, gives us a hint that ZNF146 is possibly involved in liver carcinogenesis by affecting DNA repair and cell cycle control upon induced DNA damage. / The gene ZNF146 codes for a member of the Kruppel zinc finger proteins, however ZNF146 protein is different from most members of the Kruppel zinc finger proteins subfamily. It encodes a 33 kDa protein solely composed of 10 zinc finger motifs and is devoid of any non-zinc finger regulatory domain for interactions with other proteins. ZNF146 overexpression has been reported in a number of cancers including colon cancer and pancreatic carcinoma. However, the functional role of ZNF146 overexpression in tumorigenesis is yet to be solved and not much research on how ZNF146 might be invovled in the establishment of HCC was published. / To conclude, the experimental results of this study support the hypothesis that ZNF146 overexpression may deregulating the cell division cycle and some genes differentially regulated upon over-expression of ZNF146 are related to the regulations of DNA damage response. Future research on ZNF146 can be focused on the detail regulatory pathway of ZNF146 overexpression and its interaction between the p53 tumor suppressor, DNA damage response and cell cycle regulation, and a fuller picture of how ZNF146 overexpression might induce hepatocarcinogenesis can be revealed. / Yeung, Tsz Lun. / Adviser: Miu Yee (Mary) Waye. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3329. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 287-304). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.
76

Identificação e caracterização de genes codificantes de proteínas ricas em glicina ligantes de RNA em soja (Glycine max (L.) Merril)

Poersch, Liane Balvedi January 2011 (has links)
A soja constitui uma das culturas mais importantes mundialmente, tanto social quanto economicamente. Consequentemente, informações moleculares sobre processos de desenvolvimento, bem como conhecimento detalhado das interações entre condições estressoras e a resposta da planta a fatores ambientais são necessários. A identificação e caracterização de genes que respondem a condições ambientais específicas constituem um passo inicial no entendimento dos processos adaptativos. Proteínas ricas em glicina (GRPs) são polipeptídeos contendo um grande número do aminoácido glicina em sua estrutura primária. Os genes codificantes de GRPs são regulados ao longo do desenvolvimento e regulados por auxina, ABA, frio, ferimentos, luz, ritmo circadiano, salinidade, seca, patógenos e encharcamento. Entretanto, há pouca informação sobre GRPs de plantas e seus papéis no desenvolvimento e resposta a estresses. As GRPs podem ser divididas em quatro classes (I, II, III, IV) de acordo com sua estrutura primária e presença de domínios característicos. A classe IV é composta por proteínas ligantes de RNA. Domínios adicionais permitem dividir a classe IV de GRPs em quatro subclasses (IVa, IVb, IVc, IVd). A subclasse IVc é representada por proteínas contendo um cold-schock domain (CSD) e dedos de zinco CCHC tipo retrovirais. O objetivo do presente estudo foi: (i) identificar e caracterizar os genes codificantes de classe IV de GRPs, (ii) verificar a padrão de expressão dos genes codificantes da subclasse IVc de GRPs e (iii) produzir plantas de soja transgênicas expressando o gene AtGRP2, o qual foi mostrado estar envolvido na floração e desenvolvimento da semente em Arabidopsis, e também poderia desempenhar um papel na aclimatação ao frio. Um total de 47 genes codificantes da classe IV de GRPs foi identificado no genoma da soja: 19 da subclasse IVa, sete da IVb, seis da IVc e 15 da IVd. Análises in silico indicaram uma expressão preferencial de todos os genes codificantes da subclasse IVc em tecidos em desenvolvimento. Análises de RT-qPCR revelaram que plantas jovens e maduras exibem uma expressão mais alta em folhas do que em outros órgãos, com exceção dos genes GRP2L_4/5 que tiveram expressão mais alta em sementes. GRP2L_4/5 e GRP2L_2 foram induzidos em resposta a baixas temperaturas. Sob estresse com ABA a expressão de todos os genes foi reprimida em folhas e/ou raízes, com exceção do gene GRP2L_2 que foi induzido em raízes. Em resposta a infecção com Phakopsora pachyrhizi, a expressão de GRP2L_2 e GRP2L_3 foi mais alta e precoce no genótipo suscetível quando comparada com o resistente, enquanto que a resposta de GRP2L_4/5 e GRP2L_6 foi mais tardia no genótipo resistente. Ainda, embriões somáticos secundários das cultivares Bragg, IAS-5 e BRSMG 68 Vencedora de soja foram usados para introduzir o gene AtGRP2 no genoma da soja por bombardeamento e sistema bombardeamento/Agrobacterium. Seis eventos de transformação independentes foram confirmados por PCR. No presente momento as plantas estão em desenvolvimento em frascos de vidro. No presente estudo a classe IV de GRPs em soja foi identificada e caracterizada. Este é o primeiro passo para elucidar o papel destas proteínas em plantas. / Molecular information on plant developmental process, as well as detailed knowledge of the interaction between stress conditions and plant response to environmental factors are essential for understanding the adaptive response. Glycine-Rich Proteins (GRP) have the amino acid glycine well represented in their primary structure. The genes encoding GRPs are developmentally regulated and induced by auxin, ABA, cold, wound, light, circadian rhythm, salinity, drought, pathogens, and flooding. However, there is scarce information about plant GRPs and its role on development and stress response. The GRPs can be divided into four classes (I, II, II and IV) according to their primary structure and the presence of characteristic domains. Class IV is composed by RNA-binding proteins. Additional domains permit to split class IV GRPs into four subclasses (IVa, IVb, IVc and IVd). Subclass IVc is represented by proteins containing a Cold-Shock Domain (CSD) and retroviral-like CCHC zinc fingers. The goal of the present study was: (i) to identify and characterize the genes encoding class IV GRPs, (ii) to verify the relative expression of genes encoding subclass IVc GRPs and (iii) to produce transgenic soybean plants expressing the AtGRP2 gene, which was shown to be involved in Arabidopsis flower and seed development, and can also play a role in cold acclimation. A total of 47 genes encoding class IV GRPs were found in the soybean genome: 19 from IVa, seven from IVb, six from IVc and 15 from IVd subclasses. In silico analyses indicated a preferential expression of all genes encoding subclass IVc GRPs in tissues under development. RT-qPCR analyses revealed that both young and mature plants exhibit relative higher expression of subclass IVc GRPs in leaves than in other organs, with exception of GRP2L_4/5 genes that have higher expression in seeds. The GRP2L_4/5 and GRP2L_2 were up-regulated in response to low temperatures. Under ABA stress the expression of all genes was down-regulated in leaves and roots, with exception of GRP2L_2 gene that was up-regulated in roots. In response to Phakopsora pachyrhizi infection, GRP2L_2 and GRP2L_3 expression was higher and earlier in the susceptible genotype when compared with that of the resistant one, while GRP2L_4/5 and GRP2_6 respond later in the resistant genotype. Furthermore, secondary somatic embryos of Bragg, IAS-5 and BRSMG 68 Vencedora soybean cultivars were used to introduce the AtGRP2 gene into the soybean genome by particle bombardment and bombardment/Agrobacterium system. Six independent Bragg transformation events were confirmed by PCR. In the present moment the plants are under development in glass flasks. In the present study the soybean class IV GRPs were identified and characterized. This is the first step to elucidate the role of these proteins in plants.
77

Ribonucleoprotein complexes and protein arginine methylation : a role in diseases of the central nervous sytem

Chénard, Carol Anne. January 2008 (has links)
For the past 45 years, QKI has been studied for its role in the processes of development and central nervous system myelination using the qkv mouse. The presence of a single KH domain and the recent identification of a high-affinity binding site in mRNAs, suggests that it can bind to and regulate mRNAs through processes such as stability, splicing and transport. As a member of the STAR RNA binding family of proteins the QKI isoforms may also be involved in cell signaling pathways. / QKI's involvement in all of these processes, lead us to examine both the protein partners and the mRNA targets of the QKI complex in order to identify potentially new pathways regulated by QKI. In doing so, we identified a novel direct protein-protein interaction with PABP and for the first time described the relocalization of QKI to cytoplasmic granules following oxidative stress. In addition, in vivo mRNA interaction studies were performed and allowed the identification of approximately 100 new mRNA targets in human glioblastoma cells. One of the targets identified was VEGF mRNA. / Another QKI target mRNA is MBP, a major protein component of the myelin sheath and the candidate auto-antigen in multiple sclerosis (MS). In vivo MBP is symmetrically dimethylated on a single arginine residue. To further establish the role of the methylation of MBP in myelination, a methyl-specific antibody and an adenovirus expressing a recombinant protein arginine methyltransferase 5 (PRMT5) was generated. We show that methylated MBP is found in areas of mature myelin and that overexpression of the PRTM5 blocked the differentiation of oligodendrocytes. / Taken together these datas implicate QKI for the first time in the process of human cancer angiogenesis and could explain the vascularization defects observed in some of the qkI mutant mice. In addition, arginine methylation of MBP may prove to have an important role in the process of myelination and in the pathogenesis of demyelination and the autoimmune reaction in diseases such as MS.
78

Mechanisms of T cell tolerance to the RNA-binding nuclear autoantigen human La/SS-B

Yaciuk, Jane Cherie. January 2008 (has links) (PDF)
Thesis (Ph. D.)--University of Oklahoma. / Bibliography: leaves 122-140.
79

Identificação e caracterização de genes codificantes de proteínas ricas em glicina ligantes de RNA em soja (Glycine max (L.) Merril)

Poersch, Liane Balvedi January 2011 (has links)
A soja constitui uma das culturas mais importantes mundialmente, tanto social quanto economicamente. Consequentemente, informações moleculares sobre processos de desenvolvimento, bem como conhecimento detalhado das interações entre condições estressoras e a resposta da planta a fatores ambientais são necessários. A identificação e caracterização de genes que respondem a condições ambientais específicas constituem um passo inicial no entendimento dos processos adaptativos. Proteínas ricas em glicina (GRPs) são polipeptídeos contendo um grande número do aminoácido glicina em sua estrutura primária. Os genes codificantes de GRPs são regulados ao longo do desenvolvimento e regulados por auxina, ABA, frio, ferimentos, luz, ritmo circadiano, salinidade, seca, patógenos e encharcamento. Entretanto, há pouca informação sobre GRPs de plantas e seus papéis no desenvolvimento e resposta a estresses. As GRPs podem ser divididas em quatro classes (I, II, III, IV) de acordo com sua estrutura primária e presença de domínios característicos. A classe IV é composta por proteínas ligantes de RNA. Domínios adicionais permitem dividir a classe IV de GRPs em quatro subclasses (IVa, IVb, IVc, IVd). A subclasse IVc é representada por proteínas contendo um cold-schock domain (CSD) e dedos de zinco CCHC tipo retrovirais. O objetivo do presente estudo foi: (i) identificar e caracterizar os genes codificantes de classe IV de GRPs, (ii) verificar a padrão de expressão dos genes codificantes da subclasse IVc de GRPs e (iii) produzir plantas de soja transgênicas expressando o gene AtGRP2, o qual foi mostrado estar envolvido na floração e desenvolvimento da semente em Arabidopsis, e também poderia desempenhar um papel na aclimatação ao frio. Um total de 47 genes codificantes da classe IV de GRPs foi identificado no genoma da soja: 19 da subclasse IVa, sete da IVb, seis da IVc e 15 da IVd. Análises in silico indicaram uma expressão preferencial de todos os genes codificantes da subclasse IVc em tecidos em desenvolvimento. Análises de RT-qPCR revelaram que plantas jovens e maduras exibem uma expressão mais alta em folhas do que em outros órgãos, com exceção dos genes GRP2L_4/5 que tiveram expressão mais alta em sementes. GRP2L_4/5 e GRP2L_2 foram induzidos em resposta a baixas temperaturas. Sob estresse com ABA a expressão de todos os genes foi reprimida em folhas e/ou raízes, com exceção do gene GRP2L_2 que foi induzido em raízes. Em resposta a infecção com Phakopsora pachyrhizi, a expressão de GRP2L_2 e GRP2L_3 foi mais alta e precoce no genótipo suscetível quando comparada com o resistente, enquanto que a resposta de GRP2L_4/5 e GRP2L_6 foi mais tardia no genótipo resistente. Ainda, embriões somáticos secundários das cultivares Bragg, IAS-5 e BRSMG 68 Vencedora de soja foram usados para introduzir o gene AtGRP2 no genoma da soja por bombardeamento e sistema bombardeamento/Agrobacterium. Seis eventos de transformação independentes foram confirmados por PCR. No presente momento as plantas estão em desenvolvimento em frascos de vidro. No presente estudo a classe IV de GRPs em soja foi identificada e caracterizada. Este é o primeiro passo para elucidar o papel destas proteínas em plantas. / Molecular information on plant developmental process, as well as detailed knowledge of the interaction between stress conditions and plant response to environmental factors are essential for understanding the adaptive response. Glycine-Rich Proteins (GRP) have the amino acid glycine well represented in their primary structure. The genes encoding GRPs are developmentally regulated and induced by auxin, ABA, cold, wound, light, circadian rhythm, salinity, drought, pathogens, and flooding. However, there is scarce information about plant GRPs and its role on development and stress response. The GRPs can be divided into four classes (I, II, II and IV) according to their primary structure and the presence of characteristic domains. Class IV is composed by RNA-binding proteins. Additional domains permit to split class IV GRPs into four subclasses (IVa, IVb, IVc and IVd). Subclass IVc is represented by proteins containing a Cold-Shock Domain (CSD) and retroviral-like CCHC zinc fingers. The goal of the present study was: (i) to identify and characterize the genes encoding class IV GRPs, (ii) to verify the relative expression of genes encoding subclass IVc GRPs and (iii) to produce transgenic soybean plants expressing the AtGRP2 gene, which was shown to be involved in Arabidopsis flower and seed development, and can also play a role in cold acclimation. A total of 47 genes encoding class IV GRPs were found in the soybean genome: 19 from IVa, seven from IVb, six from IVc and 15 from IVd subclasses. In silico analyses indicated a preferential expression of all genes encoding subclass IVc GRPs in tissues under development. RT-qPCR analyses revealed that both young and mature plants exhibit relative higher expression of subclass IVc GRPs in leaves than in other organs, with exception of GRP2L_4/5 genes that have higher expression in seeds. The GRP2L_4/5 and GRP2L_2 were up-regulated in response to low temperatures. Under ABA stress the expression of all genes was down-regulated in leaves and roots, with exception of GRP2L_2 gene that was up-regulated in roots. In response to Phakopsora pachyrhizi infection, GRP2L_2 and GRP2L_3 expression was higher and earlier in the susceptible genotype when compared with that of the resistant one, while GRP2L_4/5 and GRP2_6 respond later in the resistant genotype. Furthermore, secondary somatic embryos of Bragg, IAS-5 and BRSMG 68 Vencedora soybean cultivars were used to introduce the AtGRP2 gene into the soybean genome by particle bombardment and bombardment/Agrobacterium system. Six independent Bragg transformation events were confirmed by PCR. In the present moment the plants are under development in glass flasks. In the present study the soybean class IV GRPs were identified and characterized. This is the first step to elucidate the role of these proteins in plants.
80

Identificação e caracterização de genes codificantes de proteínas ricas em glicina ligantes de RNA em soja (Glycine max (L.) Merril)

Poersch, Liane Balvedi January 2011 (has links)
A soja constitui uma das culturas mais importantes mundialmente, tanto social quanto economicamente. Consequentemente, informações moleculares sobre processos de desenvolvimento, bem como conhecimento detalhado das interações entre condições estressoras e a resposta da planta a fatores ambientais são necessários. A identificação e caracterização de genes que respondem a condições ambientais específicas constituem um passo inicial no entendimento dos processos adaptativos. Proteínas ricas em glicina (GRPs) são polipeptídeos contendo um grande número do aminoácido glicina em sua estrutura primária. Os genes codificantes de GRPs são regulados ao longo do desenvolvimento e regulados por auxina, ABA, frio, ferimentos, luz, ritmo circadiano, salinidade, seca, patógenos e encharcamento. Entretanto, há pouca informação sobre GRPs de plantas e seus papéis no desenvolvimento e resposta a estresses. As GRPs podem ser divididas em quatro classes (I, II, III, IV) de acordo com sua estrutura primária e presença de domínios característicos. A classe IV é composta por proteínas ligantes de RNA. Domínios adicionais permitem dividir a classe IV de GRPs em quatro subclasses (IVa, IVb, IVc, IVd). A subclasse IVc é representada por proteínas contendo um cold-schock domain (CSD) e dedos de zinco CCHC tipo retrovirais. O objetivo do presente estudo foi: (i) identificar e caracterizar os genes codificantes de classe IV de GRPs, (ii) verificar a padrão de expressão dos genes codificantes da subclasse IVc de GRPs e (iii) produzir plantas de soja transgênicas expressando o gene AtGRP2, o qual foi mostrado estar envolvido na floração e desenvolvimento da semente em Arabidopsis, e também poderia desempenhar um papel na aclimatação ao frio. Um total de 47 genes codificantes da classe IV de GRPs foi identificado no genoma da soja: 19 da subclasse IVa, sete da IVb, seis da IVc e 15 da IVd. Análises in silico indicaram uma expressão preferencial de todos os genes codificantes da subclasse IVc em tecidos em desenvolvimento. Análises de RT-qPCR revelaram que plantas jovens e maduras exibem uma expressão mais alta em folhas do que em outros órgãos, com exceção dos genes GRP2L_4/5 que tiveram expressão mais alta em sementes. GRP2L_4/5 e GRP2L_2 foram induzidos em resposta a baixas temperaturas. Sob estresse com ABA a expressão de todos os genes foi reprimida em folhas e/ou raízes, com exceção do gene GRP2L_2 que foi induzido em raízes. Em resposta a infecção com Phakopsora pachyrhizi, a expressão de GRP2L_2 e GRP2L_3 foi mais alta e precoce no genótipo suscetível quando comparada com o resistente, enquanto que a resposta de GRP2L_4/5 e GRP2L_6 foi mais tardia no genótipo resistente. Ainda, embriões somáticos secundários das cultivares Bragg, IAS-5 e BRSMG 68 Vencedora de soja foram usados para introduzir o gene AtGRP2 no genoma da soja por bombardeamento e sistema bombardeamento/Agrobacterium. Seis eventos de transformação independentes foram confirmados por PCR. No presente momento as plantas estão em desenvolvimento em frascos de vidro. No presente estudo a classe IV de GRPs em soja foi identificada e caracterizada. Este é o primeiro passo para elucidar o papel destas proteínas em plantas. / Molecular information on plant developmental process, as well as detailed knowledge of the interaction between stress conditions and plant response to environmental factors are essential for understanding the adaptive response. Glycine-Rich Proteins (GRP) have the amino acid glycine well represented in their primary structure. The genes encoding GRPs are developmentally regulated and induced by auxin, ABA, cold, wound, light, circadian rhythm, salinity, drought, pathogens, and flooding. However, there is scarce information about plant GRPs and its role on development and stress response. The GRPs can be divided into four classes (I, II, II and IV) according to their primary structure and the presence of characteristic domains. Class IV is composed by RNA-binding proteins. Additional domains permit to split class IV GRPs into four subclasses (IVa, IVb, IVc and IVd). Subclass IVc is represented by proteins containing a Cold-Shock Domain (CSD) and retroviral-like CCHC zinc fingers. The goal of the present study was: (i) to identify and characterize the genes encoding class IV GRPs, (ii) to verify the relative expression of genes encoding subclass IVc GRPs and (iii) to produce transgenic soybean plants expressing the AtGRP2 gene, which was shown to be involved in Arabidopsis flower and seed development, and can also play a role in cold acclimation. A total of 47 genes encoding class IV GRPs were found in the soybean genome: 19 from IVa, seven from IVb, six from IVc and 15 from IVd subclasses. In silico analyses indicated a preferential expression of all genes encoding subclass IVc GRPs in tissues under development. RT-qPCR analyses revealed that both young and mature plants exhibit relative higher expression of subclass IVc GRPs in leaves than in other organs, with exception of GRP2L_4/5 genes that have higher expression in seeds. The GRP2L_4/5 and GRP2L_2 were up-regulated in response to low temperatures. Under ABA stress the expression of all genes was down-regulated in leaves and roots, with exception of GRP2L_2 gene that was up-regulated in roots. In response to Phakopsora pachyrhizi infection, GRP2L_2 and GRP2L_3 expression was higher and earlier in the susceptible genotype when compared with that of the resistant one, while GRP2L_4/5 and GRP2_6 respond later in the resistant genotype. Furthermore, secondary somatic embryos of Bragg, IAS-5 and BRSMG 68 Vencedora soybean cultivars were used to introduce the AtGRP2 gene into the soybean genome by particle bombardment and bombardment/Agrobacterium system. Six independent Bragg transformation events were confirmed by PCR. In the present moment the plants are under development in glass flasks. In the present study the soybean class IV GRPs were identified and characterized. This is the first step to elucidate the role of these proteins in plants.

Page generated in 0.1304 seconds