Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / No presente trabalho à feita a descriÃÃo de trÃs genes distintos que codificam lectinas ou proteÃnas relacionadas à lectina de Vatairea macrocarpa. As sequÃncias foram obtidas pela amplificaÃÃo de DNA genÃmico e cDNA de folhas utilizando primers semi-degenerados construÃdos a partir da informaÃÃo da sequÃncia de aminoÃcidos da lectina VML depositada no GenBank. O resultado do sequenciamento revelou a presenÃa de trÃs contigs. O contig1 corresponde à lectina VML desde que se assuma que a lectina depositada VML contenha heterogeneidades ou ambiguidades decorrentes na degradaÃÃo de Edman. A traduÃÃo dos contigs 2 e 3 mostram identidade de sequÃncia de 77% quando comparadas com VML. As sequÃncias, apesar de apresentar regiÃes conservadas, mostram diferenÃas de aminoÃcidos nos sÃtios de N-glicosilaÃÃo, sÃtios de ligaÃÃo a carboidrato e metais alÃm da presenÃa de resÃduos de cisteÃna sugerindo que tais proteÃnas podem ter outras atividades biolÃgicas. A anÃlise da sequÃncia obtida pelo 3â RACE se mostrou complementar ao contig3. Sendo assim, a sequÃncia hÃbrida contig3/contigA possui 2 resÃduos de cisteÃna alÃm de revelar diferenÃas de aminoÃcidos na regiÃo C-terminal quando alinhada com outras lectinas de leguminosas. AnÃlises filogenÃticas revelaram que os contigs observados formam um grupo monofiletico e tem alta similaridade com as lectinas de Sohora japonica e Robinia pseudoacacia, alÃm da proteÃna relacionada à lectina de Cladrastis lutea. / In this paper is made a description of three distinct genes that encode Vatairea macrocarpa lectin and related proteins. The sequences were obtained by amplification of genomic DNA and cDNA of leaves using semi - degenerate primers constructed from the information of the amino acid sequence of VML lectin deposited in GenBank. The result of sequencing rev eals the presence of three different genes, called contig 1, 2 and 3 . The VML lectin corresponds to contig1 long as one assumes that the lectin contains heterogeneities deposited VML or ambiguities arising in the Edman degradation . The translation of cont igs 2 and 3 show sequence identity of 77% compared to VML. Sequences, despite having conserved regions show differences in amino acid N - glycosylation sites, carbohydrate binding sites and metals and the presence of cysteine residues suggests that these pro teins may have other biological activities . The analysis of the sequence obtained by 3 'RACE proved complementary to contig3. Thus, the sequence contig3/contigA hybrid has two cysteine residues in addition to revealing differences in amino acid C - terminal region when aligned with other legume lectins. Phylogenetic analysis revealed that the observed contigs form a monophyletic group and has high similarity with lectins from Robinia pseudoacacia Sohora japonica and, in addition to the lectin - related protein Cladrastis lutea .

ALVES FILHO, João Garcia. Clonagem, sequenciamento e caracterização parcial dos genes relacionados à lectina de Vatairea macrocarpa. 2008. 81 f. Dissertação (Mestrado)-Universidade Federal do Ceará, Fortaleza-CE, 2008. / Submitted by Eric Santiago (erichhcl@gmail.com) on 2016-05-25T14:10:53Z No. of bitstreams: 1 2008_dis_jgalvesfilho.pdf: 1626131 bytes, checksum: fbd1ed851edb7139baaf9677e39e516a (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-07-05T20:59:57Z (GMT) No. of bitstreams: 1 2008_dis_jgalvesfilho.pdf: 1626131 bytes, checksum: fbd1ed851edb7139baaf9677e39e516a (MD5) / Made available in DSpace on 2016-07-05T20:59:57Z (GMT). No. of bitstreams: 1 2008_dis_jgalvesfilho.pdf: 1626131 bytes, checksum: fbd1ed851edb7139baaf9677e39e516a (MD5) Previous issue date: 2008 / In this paper is made a description of three distinct genes that encode Vatairea macrocarpa lectin and related proteins. The sequences were obtained by amplification of genomic DNA and cDNA of leaves using semi - degenerate primers constructed from the information of the amino acid sequence of VML lectin deposited in GenBank. The result of sequencing rev eals the presence of three different genes, called contig 1, 2 and 3 . The VML lectin corresponds to contig1 long as one assumes that the lectin contains heterogeneities deposited VML or ambiguities arising in the Edman degradation . The translation of cont igs 2 and 3 show sequence identity of 77% compared to VML. Sequences, despite having conserved regions show differences in amino acid N - glycosylation sites, carbohydrate binding sites and metals and the presence of cysteine residues suggests that these pro teins may have other biological activities . The analysis of the sequence obtained by 3 'RACE proved complementary to contig3. Thus, the sequence contig3/contigA hybrid has two cysteine residues in addition to revealing differences in amino acid C - terminal region when aligned with other legume lectins. Phylogenetic analysis revealed that the observed contigs form a monophyletic group and has high similarity with lectins from Robinia pseudoacacia Sohora japonica and, in addition to the lectin - related protein Cladrastis lutea. / No presente trabalho é feita a descrição de três genes distintos que codificam lectinas ou proteínas relacionadas à lectina de Vatairea macrocarpa. As sequências foram obtidas pela amplificação de DNA genômico e cDNA de folhas utilizando primers semi-degenerados construídos a partir da informação da sequência de aminoácidos da lectina VML depositada no GenBank. O resultado do sequenciamento revelou a presença de três contigs. O contig1 corresponde à lectina VML desde que se assuma que a lectina depositada VML contenha heterogeneidades ou ambiguidades decorrentes na degradação de Edman. A tradução dos contigs 2 e 3 mostram identidade de sequência de 77% quando comparadas com VML. As sequências, apesar de apresentar regiões conservadas, mostram diferenças de aminoácidos nos sítios de N-glicosilação, sítios de ligação a carboidrato e metais além da presença de resíduos de cisteína sugerindo que tais proteínas podem ter outras atividades biológicas. A análise da sequência obtida pelo 3’ RACE se mostrou complementar ao contig3. Sendo assim, a sequência híbrida contig3/contigA possui 2 resíduos de cisteína além de revelar diferenças de aminoácidos na região C-terminal quando alinhada com outras lectinas de leguminosas. Análises filogenéticas revelaram que os contigs observados formam um grupo monofiletico e tem alta similaridade com as lectinas de Sohora japonica e Robinia pseudoacacia, além da proteína relacionada à lectina de Cladrastis lutea.

Ma thèse s'inscrit dans l'étude des modifications du globule rouge humain induites par P. falciparum. Ces modifications qui représentent une remarquable adaptation du parasite à un environnement plus complexe qu'il n'y paraît au premier abord et expliquent sa persistance chez l'Homme sont détaillées dans une revue et un chapitre de livre dont je suis co-auteur. Mes travaux de recherche ont porté sur la caractérisation fonctionnelle des structures de Maurer, un compartiment membranaire exporté par le parasite dans le globule rouge parasitaire et directement lié à la physiopathologie du paludisme grave. J'ai contribué à la caractérisation fonctionnelle de nouvelles protéines de ces structures, codées par trois familles multigéniques sub-télomériques en cluster avec la famille Pfmc-2tm, et présentant de façon étonnante un fort degré de conservation (article 1). La diminution d'expression de ces gènes, obtenue par titration d'un facteur transcriptionnel, entraine un défaut de libération des mérozoïtes. Mon deuxième projet porte sur l'identification des modalités d'export de la protéine transmembranaire résidente des structures de Maurer PfSBP1. Mes travaux montrent que PfSBP1 est exportée sous forme soluble dans le cytoplasme érythrocytaire, en interaction avec le complexe chaperon parasitaire PfTCP1 (article 2). / Plasmodium falciparum causes the most severe forms of human malaria, a pathology associated with the erythrocytic asexual stages of the parasite. My work focused on the remodeling of the infected erythrocytes induced by P. falciparum and detailed in a review and a book chapter that I co-authored. These modifications illustrate a remarkable adaptation of P. falciparum resulting in its persistence in humans. My PhD thesis was dedicated to the functional characterization of Maurer's clefts, a membrane compartment transposed by the parasite in the cytoplasm of its host cell, and central to the export of virulence factors to the host cell surface. I have conducted two projects and contributed first to the functional characterization of novel exported protein encoded by three highly conserved multigene sub-telomeric families in cluster with the Pfmc-2tm family. Down regulation of these gene families by promoter titration impacted the release of infectious merozoites from the host cell (annex 1). My second project was dedicated to the identification of the modality of export of the resident and Maurer's clefts transmembrane protein PfSBP1. I have shown that PfSBP1 is exported as a soluble protein in the host cell cytoplasm in interaction with the parasite Thermosome complex protein 1 (PfTCP1) chaperone complex (annex 2).