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Made available in DSpace on 2016-04-28T19:58:56Z (GMT). No. of bitstreams: 1 Fernanda Marques Granato.pdf: 1246024 bytes, checksum: a72edaae43263f88c7f64c66a141f42e (MD5) Previous issue date: 2015-10-06 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This dissertation has as its aim to retrieve the concept of the nonsense genre with the purpose of testing the hypothesis of a possible dialogue between texts from two literary works of different characteristics and of different times: Viagem numa peneira, by Edward Lear (1846), and Quatro-olhos, by Renato Pompeu (1976). The studies carried out were able to ascertain that, among the analyzed texts, there are traces that make possible not only to bring the two closer together, but also to do a reinterpretation of the nonsense genre in the context of the literary contemporaneity. In order to develop this proposal, we have elected a few authors that give support to the analysis of the fictional works, such as: Aristotle (2011) and Croce (1995), on genre; Sewell (1952), Stewart (1978), Ede (1987) and Tigges (1988), on nonsense; Huizinga (2010), on game; Watt (2010) and Lukács (2009) on novel; Nikolayeva (2011), on illustration; Pignatari (2011) and Huxley (1948), on poetry; and Agamben (2009), on the contemporaneity. Departing from a historic conceptual recover, our attention is drawn to the perspective of confirmation of the hypothesis about the nonsense literary genre, its connection to the Victorian ages and the reverberations that the referred genre has brought to our contemporaneity, and specially to the novel by Renato Pompeu, leaving as credit the tendency for the fragmentation of the narrative / Esta pesquisa tem por objetivo resgatar o conceito do gênero nonsense com a finalidade de testar a hipótese de um possível diálogo entre textos de duas obras de características e épocas distintas: Viagem numa peneira, de Edward Lear (1846) e Quatro-olhos, de Renato Pompeu (1976). Os estudos realizados permitem comprovar que, entre os textos analisados, existem traços que possibilitam não apenas aproximá-los, mas também realizar uma reinterpretação do gênero nonsense no contexto da contemporaneidade literária. Para o desenvolvimento do trabalho, elegemos alguns teóricos que dão suporte à análise das obras ficcionais, são eles: Aristóteles (2011) e Croce (1995), sobre gênero; Sewell (1952), Stewart (1978), Ede (1987) e Tigges (1988), sobre nonsense; Huizinga (2010), sobre o lúdico e o jogo; Watt (2010) e Lukács (2009), sobre a teoria do romance; Nikolayeva (2011), sobre a ilustração; Pignatari (2011) e Huxley (1948), sobre a poesia; e Agamben (2009), sobre o contemporâneo. A partir de uma retomada histórico-conceitual, nossa atenção volta-se para a perspectiva de comprovação da hipótese sobre o gênero nonsense, sua ligação com a era vitoriana e as reverberações que o referido gênero traz para a nossa contemporaneidade e, em especial, para o romance de Renato Pompeu, deixando como saldo a tendência para a fragmentação narrativa

Le processus aléatoire des recombinaisons V(D)J permet d’obtenir un répertoire d’anticorps (Ac) ou immunoglobulines (Ig) hautement diversifié. En revanche, le caractère imprécis des jonctions V(D)J conduit à l’apparition de décalages du cadre de lecture dans deux tiers des cas. Ainsi, la plupart des cellules B hébergent des allèles d’Ig avec des réarrangements V(D)J non-productifs au sein de leur génome. Plusieurs études incluant celles menées au laboratoire ont montré que ces allèles non-productifs sont transcrits mais subissent une régulation post-transcriptionnelle impliquant le mécanisme de dégradation des ARNm appelé NMD « Nonsense-Mediated mRNA Decay ». Cette surveillance ARN diminue ainsi le taux d’ARNm codant pour des chaînes d’Ig tronquées. En revanche, l’impact de l’épissage alternatif des transcrits d’Ig non-productifs sur la production d’Ig aberrantes reste jusqu’ici peu exploré. L’étude de ce processus appelé NAS (« Nonsense-associated Altered Splicing »), et en particulier du phénomène de saut d’exon, présente un grand intérêt car cet épissage alternatif peut permettre la synthèse d’Ig tronquées présentant des délétions internes. Les projets développés lors de cette thèse ont révélé la toxicité des chaînes d’Ig dépourvues de domaine variable (V) dans les plasmocytes, et mis en évidence l’existence d’un nouveau point de contrôle au cours de la différenciation plasmocytaire. Ce phénomène nommé TIE-checkpoint (Truncated-Ig Exclusion) conduisant à l’élimination des plasmocytes exprimant des Ig tronquées, est la conséquence d’un saut d’exon lors de l’épissage des transcrits Ig non-productifs. Pour étudier les évènements de NAS lors de l’épissage des transcrits d’Ig dans les plasmocytes, il faut par conséquent limiter l’activation du TIE-checkpoint. A l’aide d’un modèle murin présentant un exon non-sens additionnel au locus IgH, nous avons pu analyser in vivo l’épissage alternatif par « saut d’exon » des transcrits d’Ig non-productifs. En effet, l’élimination de cet exon addtionnel aboutit à la synthèse d’une chaîne d’Ig normale et non à la production de chaînes tronquées. Cette étude a été menée dans des cellules B primaires et des plasmocytes. Les résultats obtenus ont révélé que l’hypertranscription des gènes d’Ig, qui accompagne la différenciation plasmocytaire, favorise l’épissage alternatif des transcrits d’Ig non-productifs, par un phénomène de saut d’exon. Nous avons également étudié les éventuelles connexions entre le mécanisme de NMD, impliqué dans la surveillance des ARNm, et l’UPR (« Unfolded Protein Response ») permettant de réguler l’homéostasie protéique dans les plasmocytes. De façon originale, nous avons identifié une boucle de régulation positive entre les processus de surveillance ARN (NMD) et protéique (UPR, autophagie, protéasome). La mise en évidence de cette coopération dans les plasmocytes constitue un exemple unique au vue de la littérature et, aurait pour effet de limiter la synthèse d’Ig tronquées tout en autorisant la synthèse massive d’Ig. Enfin, nous avons étudié le rôle de l’épissage des transcrits d’Ig non-codants (appelés transcrits I « germinaux ») au cours du processus de CSR « Class Switch Recombination ». Cette étude a apporté des précisions sur le rôle des sites donneurs d’épissage des exons I et révélé que la reconnaissance de ces sites d’épissage module l’intensité de la transcription de la région « switch » S adjacente, et par conséquent, son accessibilité à AID « Activation-Inducedcytidine Deaminase » lors de la CSR. / The random V(D)J recombination process contributes to the generation of a vast immunoglobulin (Ig) repertoire. However, imprecise V(D)J junctions lead to the appearance of frameshift mutations in two-third of the cases. Hence, numerous B-lineage cells retain non-productively V(D)J rearranged Ig alleles in their genome. Several studies including ours have shown that these non-productive alleles are transcribed but rapidly degraded by NMD « Nonsense-Mediated mRNA Decay », thus decreasing the level of mRNA encoding truncated Ig. However, less is known about the impact of alternative splicing on non-productive Ig transcripts, and especially « exon skipping », with regard to the production of truncated Ig with internal deletions. During my thesis, we have shown that truncated Ig chains lacking variable (V) domain exhibted toxic effects in plasma cells revealing a new « Truncated-Ig Exclusion » (TIE-) checkpoint during plasma cell differentiation. The TIE-checkpoint eliminates plasma cell-expressing truncated Ig, as a consequence of exon skipping during splicing of non-productive Igκ transcripts. However, the TIE checkpoint activation limits the analysis of NAS (« Nonsense associated Altered Splicing ») of Ig transcripts in plasma cells. Using a mouse model harboring an additional frameshift-inducing V exon at the IgH chain locus, we could analyze NAS of non-productive Ig transcripts in primary B cells and plasma cells. This study revealed that hypertranscription of Ig genes accompanying plasma cell differentiation favors alternative splicing of non-productive Ig transcripts. We also investigated potential connections between the NMD mechanism, involved in mRNA surveillance, and the UPR (« Unfolded Protein Response ») pathway that regulates protein homeostasis in plasma cells. Interestingly, we identified a positive regulatory loop between RNA (NMD) and protein (UPR, autophagy, proteasome) surveillance processes. In view of the literature, the occurrence of such cooperation is unique to plasma cells, and this should help to limit the expression of truncated Ig while allowing massive Ig synthesis. Finally, we studied other aspects of Ig RNA splicing, and investigated the role of splice donor site on non-coding « germline » I transcripts during CSR (« Class Switch Recombination »). Using dedicated mouse models, we found that the deletion of Iƴ1 splice donor site drastically decreased CSR to IgG1. Overall, this study demonstrated that the recognition of I exon donor splice site enhances transcription of « switch » regions S, facilitating their opening and the subsequent recruitment of AID « Activation-Induced cytidine Deaminase » during CSR.

Nonsense-mediated mRNA decay (NMD) specifically targets mRNAs with premature translation termination codons for rapid degradation. NMD is a highly conserved translation-dependent mRNA decay pathway, and its core Upf factors are thought to be recruited to prematurely terminating mRNP complexes, possibly through the release factors that orchestrate translation termination. Upf1 is the central regulator of NMD and recent studies have challenged the notion that this protein is specifically targeted to aberrant, nonsense-containing mRNAs. Rather, it has been proposed that Upf1 binds to most mRNAs in a translation-independent manner. In this thesis, I investigated the nature of Upf1 association with its substrates in the yeast Saccharomyces cerevisiae. Using biochemical and genetic approaches, the basis for Upf1 interaction with ribosomes was evaluated to determine the specificity of Upf1 association with ribosomes, and the extent to which such binding is dependent on prior association of Upf1’s interacting partners. I discovered that Upf1 is specifically associated with Rps26 of the 40S ribosomal subunit, and that this association requires the N-terminal Upf1 CH domain. In addition, using selective ribosome profiling, I investigated when during translation Upf1 associates with ribosomes and showed that Upf1 binding was not limited to polyribosomes that were engaged in translating NMD substrate mRNAs. Rather, Upf1 associated with translating ribosomes on most mRNAs, binding preferentially as ribosomes approached the 3’ ends of open reading frames. Collectively, these studies provide new mechanistic insights into NMD and the dynamics of Upf1 during translation.

Made available in DSpace on 2016-04-28T19:58:50Z (GMT). No. of bitstreams: 1 Selma Simoes.pdf: 2824155 bytes, checksum: 29fcc1f7855f8f660b27079ecf25b30d (MD5) Previous issue date: 2013-04-29 / The aim of this dissertation is to study nonsense in dialogue incorporated in the narrative process, in Lewis Carroll s masterpieces, Alice s Adventures in Wonderland and Through the Looking-Glass, with the purpose to observe the effects of the lack of coherent meaning in writing and rewriting through the characterization art illustration. In reading those works we verified how two ways of narrating/presenting were brought together by Nineteenth-century through the mediation of the logic of nonsense. Such a connection introduced certain markers in the discourse, and these work out a transposition form the logical to the illogical, in both verbal and non-verbal communication. Such an operation produced events of language, of inversion, and of semantic meaninglessness or emptying of meaning, which were interpreted by the readers- illustrators, as modes of artistic exercise by Lewis Carroll, John Tenniel modern and contemporary illustrators that make up the focus of our texts in the analysis and interpretation. In order to understand mediation of the nonsensical word and recognize it in the modes of creative interpretation exercise, we propose three hypothesis which were expectedly demonstrated and interpreted by resorting the multiple theoretical supports. The first demands the correlation and equivalence between the verbal and visual enunciates counterpoising each other in the communication process. The second indicates the perceptive, semantic and artistic markers of differences present in verbal and visual enunciates as indexing units dialogue replies. The last situates word and image as mediated by nonsense devoid of time-frontiers, pushing Carroll s narratives towards a polysemy of values and senses by means of the art to characterize illustration as works of an infinite significance / O objetivo desta dissertação é o estudo da figura do nonsense nas obras Alice no País das Maravilhas(1865) e Através do Espelho(1872),de Lewis Carroll,integrada ao diálogo no processo do narrar,cuja finalidade é observar seus efeitos sobre a escritura e reescritura pela via da arte de caracterização da ilustração.Ao ler e analisar essas obras,identificamos que a aproximação das duas formas de narrar/mostrar dessas narrativas do século XIX,pela mediação da lógica do nonsense,fazem uso de certos marcadores no discurso,e estes executam um trabalho de transposição da lógica para a não lógica,tanto na comunicação verbal quanto não verbal.Esta transmutação produz eventos de linguagem,de inversão,esvaziamento e negação semânticos,trazidos pelos leitores e ilustradores dessas narrativas como modos de exercício do trabalho artístico realizado por Lewis Carroll,John Tenniel,ilustradores modernos e contemporâneos,que constituem o nosso corpus textual de análise e interpretação.Para atender a essa mediação da palavra nonsensical e poder reconhecê-la nos modos do exercício da interpretação criativa,propomos três hipóteses,que foram demonstradas e interpretadas sob múltiplos suportes teóricos:a primeira,prevê a correlação e a equivalência entre o enunciado verbal e o visual que se contrapõem no processo de comunicação;a segunda aponta para os marcadores das diferenças(semânticas,perceptivas e artísticas)presentes no enunciado verbal e visual como unidades de réplicas do diálogo;a terceira situa o diálogo palavra e imagem mediado pelo nonsense sem fronteiras temporais,impulsionando as narrativas de Carroll à polissemia de valores e sentido pela via da arte de caracterizar da ilustração como obras de significância infinita

O complexo da esclerose tuberosa (TSC) é um transtorno genético, sistêmico, com expressividade variável e herança autossômica dominante. Clinicamente manifesta-se devido ao desenvolvimento de hamartomas e hamártias em diferentes tecidos, principalmente no cérebro, rins, coração, pele e pulmões, podendo causar disfunção do órgão. Mutação em um de dois genes supressores tumorais, TSC1 ou TSC2, são responsáveis pelo TSC. Os genes TSC1 e TSC2 codificam para hamartina e tuberina, respectivamente. Ambas as proteínas interagem fisicamente formando um complexo citosólico que inibe mTOR (mammalian target of rapamycin). Testes moleculares para TSC1 e TSC2 são úteis para auxiliar no diagnóstico de casos clínicos difíceis, em aconselhamento genético e estudos de associação genótipo-fenotípica, além de permitirem a caracterização molecular de mecanismos patogenéticos da formação dos hamartomas e análises funcionais de seus produtos gênicos. Apesar de o diagnóstico de TSC ser basicamente clínico, a partir da revisão de seus critérios em 2012 por um grupo de especialistas, o achado de uma mutação em TSC1 ou TSC2 passou a ser considerado suficiente para o diagnóstico definitivo da doença. O estudo apresentado aqui é parte de um projeto em andamento para estabelecer condições ao desenvolvimento de análise de mutações causadoras de TSC nos genes TSC1 e TSC2. Nosso objetivo neste trabalho foi avaliar por sequenciamento de Sanger o DNA genômico de 28 pacientes brasileiros com diagnóstico definitivo de TSC, procedentes dos estados de São Paulo ou Paraná, tendo como alvo a sequência codificadora do gene TSC1, parte de seus segmentos intrônicos, o promotor basal, bem como o segmento imediatamente a 5\' deste. Sete pacientes (25%) apresentaram mutações de sentido trocado (nonsense) ou com deslocamento da leitura à tradução (frameshift) no gene TSC1. Entre 31 outras variantes de DNA encontradas, 23 são polimorfismos conhecidos e oito apresentaram frequência inferior a 1%, como verificado in silico entre mais de mil sequências de genomas humanos. Entre essas oito variantes de DNA novas ou raras, quatro foram detectadas em pacientes para os quais uma mutação patogênica havia sido identificada e, por isso, foram reclassificadas como polimorfismos. Duas e uma variantes de DNA do mesmo paciente flanqueavam um sítio de ligação em potencial para um fator de transcrição específico, a 5\' do promotor basal de TSC1. Por fim, uma nova variante de DNA na região não codificadora do éxon 2 do gene TSC1 foi predita com potencial para alterar um elemento candidato a acentuador de splicing. Em resumo, como observado em estudos anteriores, descrevemos aqui 25% dos pacientes com TSC apresentando mutações patogênicas na sequência codificadora do gene TSC1. Nossos dados mostraram quatro novas variantes de DNA em regiões potencialmente reguladoras da expressão do gene TSC1, que podem revelar-se como mutações patogênicas e, portanto, necessitam ser testadas experimentalmente / Tuberous sclerosis complex (TSC) is a multisystem disorder, with variable expression and autosomal dominant inheritance. Clinically it is due to hamartia and hamartoma development in different tissues, notably in the brain, kidneys, heart, skin and lungs, causing organ dysfunction. Mutations in either tumor suppressor gene, TSC1 or TSC2, are responsible for TSC. TSC1 and TSC2 genes code for hamartin and tuberin, respectively. Both proteins physically interact forming a cytosolic complex that inhibits the mammalian target of rapamycin (mTOR). TSC1 and TSC2 molecular testing has been useful in diagnosing clinically challenging cases, in genetic counseling and genotype/phenotype association studies, besides evaluation of the molecular basis of hamartoma formation and functional analyses of both gene products. Although TSC diagnosis is basically clinical, since the 2012 specialist panel review the finding of a TSC1 or TSC2 mutation has been considered sufficient for the definite diagnosis of the disease. The results presented here are part of an ongoing project to establish conditions for TSC1 and TSC2 mutation studies. Our first aim is to evaluate by Sanger sequencing TSC1 coding sequence, and an average of 132 base pairs of intronic regions next to exon boundaries from TSC patients, in addition to the gene core promoter. We present preliminary results of a sample of 28 patients with definite TSC diagnosis, from São Paulo and Paraná states. Seven patients (25%) displayed TSC1 nonsense or frameshift mutations. Among 31 other DNA variants identified, 23 were known polymorphisms, and eight had frequencies below 1% as verified in silico among more than a thousand human genomes. Out of eight novel or rare DNA variants, four were detected in patients for whom a pathogenic mutation had been found. Two and one additional DNA point variants from the same patient flanked a putative transcription factor binding site. Finally, a novel DNA variant residing in the TSC1 noncoding exon 2 was predicted to change the sequence potential to behave as a splicing enhancer. In summary, similar to previous studies, we describe 25% of TSC patients with mutations in the TSC1 coding sequence. Differently from other reports, our data disclose four novel DNA variants in TSC1 potentially regulatory regions that are likely to unravel novel pathogenic mutations, and thus need to be experimentally tested

Hintergrund: Der Insulin-like growth factor receptor (IGF1R) spielt eine zentrale Rolle bei Wachstumsprozessen. Heterozygote IGF1R-Mutationen führen durch eine partielle IGF1-Resistenz zu Kleinwuchs. Methoden: Auxologische und endokrinologische Daten des Patienten wurden erhoben. Anhand von Fibroblasten wurde die IGF1R-Deletion charakterisiert und die Auswirkungen auf die mRNA- und Protein-Expression sowie die Signaltransduktion untersucht. Ergebnisse: Der Junge, der eine heterozygote Exon 6 Deletion im IGF1R – durch Alu-Rekombination verursacht – und eine heterozygote SHOX-Variante (p.Met240Ile) in seinem Genom vereint, kam ‚appropriate for gestational age‘ zur Welt, entwickelte aber postnatal eine Wachstumsretardierung. Die Endokrinologischen Daten waren unauffällig. Der Patient zeigt keine Stigmata, die bei anderen IGF1- oder SHOX-Mutationsträgern beschrieben wurden. Durch Nonsense-Mediated mRNA Decay kommt es zu einer Dosisreduktion der IGF1-Rezeptoren und einer entsprechenden verminderten Aktivierung der Rezeptoren, nicht aber des Signalwegs. Zusammenfassung: Der Patient trägt eine bisher unbeschrieben heterozygote IGF1R-Deletion, die zu Kleinwuchs führt. Ursächlich dafür ist eine durch die Mutation verursachte Dosisreduktion der IGF1-Rezeptoren.

Cette thèse consiste en l’analyse du jeu dans les deux œuvres littéraires majeures de Lewis Carroll, Alice’s Adventures in Wonderland et Through the Looking-Glass, œuvres qui accordent la part belle au jeu, tant au niveau diégétique, narratologique que stylistique et linguistique. Il en ressort qu’une tension entre liberté et règle (entre paidia, l’expression impulsive d’un instinct de jeu, et ludus, le besoin de créer des règles et de s’y plier, pour utiliser les termes de Caillois) traverse les Alice. L’étude des jeux et jouets, des macro- et micro-structures, du style et des intertextes, permet d’affirmer que ces deux volumes sont résolument ludiques, car ils reposent sur une légaliberté, une liberté dans et par une légalité (Duflo). Ils jouent avec et contre les attentes, la langue et les connaissances du lecteur de sens commun. Toutefois, dans le même temps, le champ d’action du lecteur virtuel est considérablement restreint : ses facultés d’idéation sont orientées par l’imbrication du texte et des illustrations, et sa participation à la construction du texte carrollien se borne à compléter les « blancs » textuels, tel que Eco les définit, c’est-à-dire à faire ressurgir les déjà-dits qui ont été effacés. Les œuvres carrolliennes sont ainsi caractérisées par ce paradoxe : alors que la diégèse et l’économie textuelle semblent promouvoir le jeu, le rôle du Lecteur Modèle prévu par le texte est extrêmement réduit. Cependant, il est possible pour le lecteur réel, interpellé par le texte carrollien et son Auteur, d’endosser le rôle de Lecteur Imposteur, de les contre-interpeller, et de devenir, par ce processus de subjectivation, un lecteur pleinement joueur. Il s’avère alors que la légaliberté permet non seulement de saisir l’esthétique du jeu, mais aussi la formation des sujets (personnages, auteurs, lecteurs) à l’œuvre dans les Alice. / This thesis consists in an analysis of play and game(s) in Lewis Carroll’s two major literary works, Alice’s Adventures in Wonderland and Through the Looking-Glass, in which diegetic, narratological, stylistic and linguistic games play a significant part. It shows that a tension is at work throughout the Alice books, between freedom and rules (between paidia, the impulsive manifestation of a play instinct, and ludus, the need to invent rules and to abide by them, to state this in Caillois’s terms). The study of games and toys, of the macro- and micro-structures, of the style and the intertexts reveals the playfulness of these texts, as they rely on “legafreedom”, freedom in, and made possible by legality (Duflo). They play with and against the reader’s commonsensical expectations, language and knowledge. Yet, at the same time, the virtual reader’s playing field is considerably limited: his or her faculties of ideation are directed by the interweaving of text and illustrations, and his or her participation (involvement?) in constructing the Carrollian text is restricted to filling in the textual “blanks,” as defined by Eco, i.e. making the “already said” that has been erased reappear. Carroll’s works are, therefore, characterized by this paradoxical idea: while the diegesis and the textual economy seem to promote play, the role of the Model Reader as mapped out by the text is extremely circumscribed. However, the real reader, interpellated by the Carrollian text and its Author, can take on the role of the Impostor Reader, counter-interpellate them, and become, thanks to this process of subjectification, a consummate playing reader. As such, “legafreedom” makes it possible to understand not only the aesthetics of play, but also the formation of subjects (characters, authors, readers) in the Alice books.

nÃo hà / Esta pesquisa busca analisar os elementos utilizados por Tim Burton para representar a Era Vitoriana atravÃs do figurino, meio de transporte e decoraÃÃo. Trabalharemos tambÃm com a traduÃÃo em imagens do nonsense em sua adaptaÃÃo fÃlmica Alice in Wonderland (2010). Nessa perspectiva, o presente trabalho resulta de um estudo comparativo entre o filme e a obra na qual se baseia, o cÃnone da literatura infantil Aliceâs Adventures in Wonderland de Lewis Carroll (1865). Para tanto, faremos um cotejo entre as duas obras à luz das teorias de Elizabeth Sewell (2015), Wim Tigges, (1988) e Susan Stewart (1980) que alicerÃam o gÃnero literÃrio nonsense. LanÃaremos mÃo da versÃo literÃria Lewis Carroll: The Complete Fully Illustrated Works de 1995 da editora Gramercy, contendo toda a obra de Lewis Carroll com ilustraÃÃes originais. Isso, a fim de viabilizar e facilitar a comparaÃÃo com as figuras presentes no texto literÃrio, quando necessÃrio, para investigar se houve aproximaÃÃo na representaÃÃo visual feita pelo diretor.

O complexo da esclerose tuberosa (TSC) é um transtorno genético, sistêmico, com expressividade variável e herança autossômica dominante. Clinicamente manifesta-se devido ao desenvolvimento de hamartomas e hamártias em diferentes tecidos, principalmente no cérebro, rins, coração, pele e pulmões, podendo causar disfunção do órgão. Mutação em um de dois genes supressores tumorais, TSC1 ou TSC2, são responsáveis pelo TSC. Os genes TSC1 e TSC2 codificam para hamartina e tuberina, respectivamente. Ambas as proteínas interagem fisicamente formando um complexo citosólico que inibe mTOR (mammalian target of rapamycin). Testes moleculares para TSC1 e TSC2 são úteis para auxiliar no diagnóstico de casos clínicos difíceis, em aconselhamento genético e estudos de associação genótipo-fenotípica, além de permitirem a caracterização molecular de mecanismos patogenéticos da formação dos hamartomas e análises funcionais de seus produtos gênicos. Apesar de o diagnóstico de TSC ser basicamente clínico, a partir da revisão de seus critérios em 2012 por um grupo de especialistas, o achado de uma mutação em TSC1 ou TSC2 passou a ser considerado suficiente para o diagnóstico definitivo da doença. O estudo apresentado aqui é parte de um projeto em andamento para estabelecer condições ao desenvolvimento de análise de mutações causadoras de TSC nos genes TSC1 e TSC2. Nosso objetivo neste trabalho foi avaliar por sequenciamento de Sanger o DNA genômico de 28 pacientes brasileiros com diagnóstico definitivo de TSC, procedentes dos estados de São Paulo ou Paraná, tendo como alvo a sequência codificadora do gene TSC1, parte de seus segmentos intrônicos, o promotor basal, bem como o segmento imediatamente a 5\' deste. Sete pacientes (25%) apresentaram mutações de sentido trocado (nonsense) ou com deslocamento da leitura à tradução (frameshift) no gene TSC1. Entre 31 outras variantes de DNA encontradas, 23 são polimorfismos conhecidos e oito apresentaram frequência inferior a 1%, como verificado in silico entre mais de mil sequências de genomas humanos. Entre essas oito variantes de DNA novas ou raras, quatro foram detectadas em pacientes para os quais uma mutação patogênica havia sido identificada e, por isso, foram reclassificadas como polimorfismos. Duas e uma variantes de DNA do mesmo paciente flanqueavam um sítio de ligação em potencial para um fator de transcrição específico, a 5\' do promotor basal de TSC1. Por fim, uma nova variante de DNA na região não codificadora do éxon 2 do gene TSC1 foi predita com potencial para alterar um elemento candidato a acentuador de splicing. Em resumo, como observado em estudos anteriores, descrevemos aqui 25% dos pacientes com TSC apresentando mutações patogênicas na sequência codificadora do gene TSC1. Nossos dados mostraram quatro novas variantes de DNA em regiões potencialmente reguladoras da expressão do gene TSC1, que podem revelar-se como mutações patogênicas e, portanto, necessitam ser testadas experimentalmente / Tuberous sclerosis complex (TSC) is a multisystem disorder, with variable expression and autosomal dominant inheritance. Clinically it is due to hamartia and hamartoma development in different tissues, notably in the brain, kidneys, heart, skin and lungs, causing organ dysfunction. Mutations in either tumor suppressor gene, TSC1 or TSC2, are responsible for TSC. TSC1 and TSC2 genes code for hamartin and tuberin, respectively. Both proteins physically interact forming a cytosolic complex that inhibits the mammalian target of rapamycin (mTOR). TSC1 and TSC2 molecular testing has been useful in diagnosing clinically challenging cases, in genetic counseling and genotype/phenotype association studies, besides evaluation of the molecular basis of hamartoma formation and functional analyses of both gene products. Although TSC diagnosis is basically clinical, since the 2012 specialist panel review the finding of a TSC1 or TSC2 mutation has been considered sufficient for the definite diagnosis of the disease. The results presented here are part of an ongoing project to establish conditions for TSC1 and TSC2 mutation studies. Our first aim is to evaluate by Sanger sequencing TSC1 coding sequence, and an average of 132 base pairs of intronic regions next to exon boundaries from TSC patients, in addition to the gene core promoter. We present preliminary results of a sample of 28 patients with definite TSC diagnosis, from São Paulo and Paraná states. Seven patients (25%) displayed TSC1 nonsense or frameshift mutations. Among 31 other DNA variants identified, 23 were known polymorphisms, and eight had frequencies below 1% as verified in silico among more than a thousand human genomes. Out of eight novel or rare DNA variants, four were detected in patients for whom a pathogenic mutation had been found. Two and one additional DNA point variants from the same patient flanked a putative transcription factor binding site. Finally, a novel DNA variant residing in the TSC1 noncoding exon 2 was predicted to change the sequence potential to behave as a splicing enhancer. In summary, similar to previous studies, we describe 25% of TSC patients with mutations in the TSC1 coding sequence. Differently from other reports, our data disclose four novel DNA variants in TSC1 potentially regulatory regions that are likely to unravel novel pathogenic mutations, and thus need to be experimentally tested