Publications by authors named "Marina P Nobrega"

7 Publications

  • Page 1 of 1

Evidence for the role of calcineurin in morphogenesis and calcium homeostasis during mycelium-to-yeast dimorphism of Paracoccidioides brasiliensis.

Eukaryot Cell 2008 Oct 5;7(10):1856-64. Epub 2008 Sep 5.

Instituto de Pesquisa e Desenvolvimento, Universidade do Vale do Paraiba, Urbanova, Sao Paulo, Brazil.

Paracoccidioides brasiliensis is a dimorphic fungus that causes paracoccidioidomycosis, the most prevalent human deep mycosis in Latin America. The dimorphic transition from mycelium to yeast (M-Y) is triggered by a temperature shift from 25 degrees C to 37 degrees C and is critical for pathogenicity. Intracellular Ca(2+) levels increased in hyphae immediately after temperature-induced dimorphism. The chelation of Ca(2+) with extracellular (EGTA) or intracellular (BAPTA) calcium chelators inhibited temperature-induced dimorphism, whereas the addition of extracellular Ca(2+) accelerated dimorphism. The calcineurin inhibitor cyclosporine A (CsA), but not tacrolimus (FK506), effectively decreased cell growth, halted the M-Y transition that is associated with virulence, and caused aberrant growth morphologies for all forms of P. brasiliensis. The difference between CsA and FK506 was ascribed by the higher levels of cyclophilins contrasted to FKBPs, the intracellular drug targets required for calcineurin suppression. Chronic exposure to CsA abolished intracellular Ca(2+) homeostasis and decreased mRNA transcription of the CCH1 gene for the plasma membrane Ca(2+) channel in yeast-form cells. CsA had no detectable effect on multidrug resistance efflux pumps, while the effect of FK506 on rhodamine excretion was not correlated with the transition to yeast form. In this study, we present evidence that Ca(2+)/calmodulin-dependent phosphatase calcineurin controls hyphal and yeast morphology, M-Y dimorphism, growth, and Ca(2+) homeostasis in P. brasiliensis and that CsA is an effective chemical block for thermodimorphism in this organism. The effects of calcineurin inhibitors on P. brasiliensis reinforce the therapeutic potential of these drugs in a combinatory approach with antifungal drugs to treat endemic paracoccidioidomycosis.
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http://dx.doi.org/10.1128/EC.00110-08DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2568058PMC
October 2008

Characterization of Paracoccidioides brasiliensis COX9, COX12, and COX16 respiratory genes.

Mycol Res 2008 Dec 11;112(Pt 12):1414-20. Epub 2008 Jul 11.

Programa de Microbiologia, Instituto de Biociências, Universidade de São Paulo, Av. Prof. Lineu Prestes, 1374, 05508-900, São Paulo, SP, Brazil.

Paracoccidioides brasiliensis is a thermo-dimorphic fungus that is the causative agent of paracoccidioidomyicosis (PCM), a human systemic granulomatous mycosis found in Latin America. Dimorphic transition from mycelium to yeast is required for establishing pathogenicity. Dimorphism is marked by changes in mitochondrial physiology, including modulation of respiration rate. In this work, we present the identification of three P. brasiliensis nuclear genes PbCOX9, PbCOX12, and PbCOX16 that code for structural subunits and a putative assembly facilitator (PbCOX16) of the mitochondrial cytochrome c oxidase (COX), the terminal enzyme complex of the respiratory chain. We measured their expression pattern during the dimorphic transition from mycelium to yeast and back by real-time reverse transcription quantitative polymerase chain reaction (real-time RT-qPCR). Our results show that messages from these genes increase during the mycelium to yeast transition and decrease during the opposite conversion. This result supports active mitochondrial participation in the transition. Heterologous complementation of the corresponding Saccharomyces cerevisiae null mutant with the PbCOX9 gene was successfully obtained.
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http://dx.doi.org/10.1016/j.mycres.2008.07.001DOI Listing
December 2008

The complete DNA sequence of the mitochondrial genome of the dermatophyte fungus Epidermophyton floccosum.

Curr Genet 2006 May 1;49(5):302-8. Epub 2006 Feb 1.

Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, Av. Prof. Lineu Prestes, 1374, Ed. Biomédicas II, Cidade Universitária, CEP 05508-900, São Paulo, Brazil.

We report here the complete nucleotide sequence of the 30.9-kb mitochondrial genome of the dermatophyte fungus Epidermophyton floccosum. All genes are encoded on the same DNA strand and include seven subunits of the reduced nicotinamide adenine dinucleotide ubiquinone oxireductase (nad1, nad2, nad3, nad4, nad4L, nad5, and nad6), three subunits of cytochrome oxidase (cox1, cox2, and cox3), apocytochrome b (cob), three subunits of ATP synthase (atp6, atp8, and atp9), the small and large ribosomal RNAs (rns and rnl), and 25 tRNAs. A ribosomal protein gene (rps5) is present as an intronic ORF in the large ribosomal subunit. The genes coding for cob and cox1 carry one intron and nad5 carries two introns with ORFs. The mtDNA of E. floccosum has the same gene order as Trichophyton rubrum mtDNA, with the exception of some tRNA genes. Maximum likelihood phylogenetic analysis confirms T. rubrum as a close relative of E. floccosum. This is the first complete mitochondrial sequence of a species of the order Onygenales. This sequence is available under GenBank accession number AY916130.
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http://dx.doi.org/10.1007/s00294-006-0057-2DOI Listing
May 2006

A transcript finishing initiative for closing gaps in the human transcriptome.

Authors:
Mari Cleide Sogayar Anamaria A Camargo Fabiana Bettoni Dirce Maria Carraro Lilian C Pires Raphael B Parmigiani Elisa N Ferreira Eloísa de Sá Moreira Maria do Rosário D de O Latorre Andrew J G Simpson Luciana Oliveira Cruz Theri Leica Degaki Fernanda Festa Katlin B Massirer Mari C Sogayar Fernando Camargo Filho Luiz Paulo Camargo Marco A V Cunha Sandro J De Souza Milton Faria Silvana Giuliatti Leonardo Kopp Paulo S L de Oliveira Paulo B Paiva Anderson A Pereira Daniel G Pinheiro Renato D Puga Jorge Estefano S de Souza Dulcineia M Albuquerque Luís E C Andrade Gilson S Baia Marcelo R S Briones Ana M S Cavaleiro-Luna Janete M Cerutti Fernando F Costa Eugenia Costanzi-Strauss Enilza M Espreafico Adriana C Ferrasi Emer S Ferro Maria A H Z Fortes Joelma R F Furchi Daniel Giannella-Neto Gustavo H Goldman Maria H S Goldman Arthur Gruber Gustavo S Guimarães Christine Hackel Flavio Henrique-Silva Edna T Kimura Suzana G Leoni Cláudia Macedo Bettina Malnic Carina V Manzini B Suely K N Marie Nilce M Martinez-Rossi Marcelo Menossi Elisabete C Miracca Maria A Nagai Francisco G Nobrega Marina P Nobrega Sueli M Oba-Shinjo Márika K Oliveira Guilherme M Orabona Audrey Y Otsuka Maria L Paço-Larson Beatriz M C Paixão Jose R C Pandolfi Maria I M C Pardini Maria R Passos Bueno Geraldo A S Passos Joao B Pesquero Juliana G Pessoa Paula Rahal Cláudia A Rainho Caroline P Reis Tatiana I Ricca Vanderlei Rodrigues Silvia R Rogatto Camila M Romano Janaína G Romeiro Antonio Rossi Renata G Sá Magaly M Sales Simone C Sant'Anna Patrícia L Santarosa Fernando Segato Wilson A Silva Ismael D C G Silva Neusa P Silva Andrea Soares-Costa Maria F Sonati Bryan E Strauss Eloiza H Tajara Sandro R Valentini Fabiola E Villanova Laura S Ward Dalila L Zanette

Genome Res 2004 Jul 14;14(7):1413-23. Epub 2004 Jun 14.

We report the results of a transcript finishing initiative, undertaken for the purpose of identifying and characterizing novel human transcripts, in which RT-PCR was used to bridge gaps between paired EST clusters, mapped against the genomic sequence. Each pair of EST clusters selected for experimental validation was designated a transcript finishing unit (TFU). A total of 489 TFUs were selected for validation, and an overall efficiency of 43.1% was achieved. We generated a total of 59,975 bp of transcribed sequences organized into 432 exons, contributing to the definition of the structure of 211 human transcripts. The structure of several transcripts reported here was confirmed during the course of this project, through the generation of their corresponding full-length cDNA sequences. Nevertheless, for 21% of the validated TFUs, a full-length cDNA sequence is not yet available in public databases, and the structure of 69.2% of these TFUs was not correctly predicted by computer programs. The TF strategy provides a significant contribution to the definition of the complete catalog of human genes and transcripts, because it appears to be particularly useful for identification of low abundance transcripts expressed in a restricted set of tissues as well as for the delineation of gene boundaries and alternatively spliced isoforms.
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http://dx.doi.org/10.1101/gr.2111304DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC442158PMC
July 2004

Analysis and functional annotation of an expressed sequence tag collection for tropical crop sugarcane.

Genome Res 2003 Dec 12;13(12):2725-35. Epub 2003 Nov 12.

Centro de Biologia Molecular e Engenharia Genética, Instituto da Computação, Universidade Estadual de Campinas, 13083-970 Campinas-SP, Brazil.

To contribute to our understanding of the genome complexity of sugarcane, we undertook a large-scale expressed sequence tag (EST) program. More than 260,000 cDNA clones were partially sequenced from 26 standard cDNA libraries generated from different sugarcane tissues. After the processing of the sequences, 237,954 high-quality ESTs were identified. These ESTs were assembled into 43,141 putative transcripts. Of the assembled sequences, 35.6% presented no matches with existing sequences in public databases. A global analysis of the whole SUCEST data set indicated that 14,409 assembled sequences (33% of the total) contained at least one cDNA clone with a full-length insert. Annotation of the 43,141 assembled sequences associated almost 50% of the putative identified sugarcane genes with protein metabolism, cellular communication/signal transduction, bioenergetics, and stress responses. Inspection of the translated assembled sequences for conserved protein domains revealed 40,821 amino acid sequences with 1415 Pfam domains. Reassembling the consensus sequences of the 43,141 transcripts revealed a 22% redundancy in the first assembling. This indicated that possibly 33,620 unique genes had been identified and indicated that >90% of the sugarcane expressed genes were tagged.
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http://dx.doi.org/10.1101/gr.1532103DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC403815PMC
December 2003

The generation and utilization of a cancer-oriented representation of the human transcriptome by using expressed sequence tags.

Authors:
Helena Brentani Otávia L Caballero Anamaria A Camargo Aline M da Silva Wilson Araújo da Silva Emmanuel Dias Neto Marco Grivet Arthur Gruber Pedro Edson Moreira Guimaraes Winston Hide Christian Iseli C Victor Jongeneel Janet Kelso Maria Aparecida Nagai Elida Paula Benquique Ojopi Elisson C Osorio Eduardo M R Reis Gregory J Riggins Andrew John George Simpson Sandro de Souza Brian J Stevenson Robert L Strausberg Eloiza H Tajara Sergio Verjovski-Almeida Marcio Luis Acencio Mário Henrique Bengtson Fabiana Bettoni Walter F Bodmer Marcelo R S Briones Luiz Paulo Camargo Webster Cavenee Janete M Cerutti Luis Eduardo Coelho Andrade Paulo César Costa dos Santos Maria Cristina Ramos Costa Israel Tojal da Silva Marcos Roberto H Estécio Karine Sa Ferreira Frank B Furnari Milton Faria Pedro A F Galante Gustavo S Guimaraes Adriano Jesus Holanda Edna Teruko Kimura Maarten R Leerkes Xin Lu Rui M B Maciel Elizabeth A L Martins Katlin Brauer Massirer Analy S A Melo Carlos Alberto Mestriner Elisabete Cristina Miracca Leandro Lorenco Miranda Francisco G Nobrega Paulo S Oliveira Apua C M Paquola José Rodrigo C Pandolfi Maria Ines de Moura Campos Pardini Fabio Passetti John Quackenbush Beatriz Schnabel Mari Cleide Sogayar Jorge E Souza Sandro R Valentini Andre C Zaiats Elisabete Jorge Amaral Liliane A T Arnaldi Amelia Goes de Araújo Simone Aparecida de Bessa David C Bicknell Maria Eugenia Ribeiro de Camaro Dirce Maria Carraro Helaine Carrer Alex F Carvalho Christian Colin Fernando Costa Cyntia Curcio Ismael Dale Cotrim Guerreiro da Silva Neusa Pereira da Silva Márcia Dellamano Hamza El-Dorry Enilza Maria Espreafico Ari José Scattone Ferreira Cristiane Ayres Ferreira Maria Angela H Z Fortes Angelita Habr Gama Daniel Giannella-Neto Maria Lúcia C C Giannella Ricardo R Giorgi Gustavo Henrique Goldman Maria Helena S Goldman Christine Hackel Paulo Lee Ho Elza Myiuki Kimura Luiz Paulo Kowalski Jose E Krieger Luciana C C Leite Ademar Lopes Ana Mercedes S C Luna Alan Mackay Suely Kazue Nagahashi Mari Adriana Aparecida Marques Waleska K Martins André Montagnini Mario Mourão Neto Ana Lucia T O Nascimento A Munro Neville Marina P Nobrega Mike J O'Hare Audrey Yumi Otsuka Anna Izabel Ruas de Melo Maria Luisa Paco-Larson Gonçalo Guimarães Pereira Neusa Pereira da Silva Joao Bosco Pesquero Juliana Gilbert Pessoa Paula Rahal Claudia Aparecida Rainho Vanderlei Rodrigues Silvia Regina Rogatto Camila Malta Romano Janaina Gusmao Romeiro Benedito Mauro Rossi Monica Rusticci Renata Guerra de Sá Simone Cristina Sant' Anna Miriam L Sarmazo Teresa Cristina de Lima E Silva Fernando Augusto Soares Maria de Fátima Sonati Josane de Freitas Sousa Diana Queiroz Valéria Valente André Luiz Vettore Fabiola Elizabeth Villanova Marco Antonio Zago Heloisa Zalcberg

Proc Natl Acad Sci U S A 2003 Nov 30;100(23):13418-23. Epub 2003 Oct 30.

Laboratorio de Genética Molecular do Cancer, Departmento de Radiologia, Universidade de São Paulo, Travessa da Rua Dr. Ovídeo Pires de Campos S/N, 4deg, Brazil.

Whereas genome sequencing defines the genetic potential of an organism, transcript sequencing defines the utilization of this potential and links the genome with most areas of biology. To exploit the information within the human genome in the fight against cancer, we have deposited some two million expressed sequence tags (ESTs) from human tumors and their corresponding normal tissues in the public databases. The data currently define approximately 23,500 genes, of which only approximately 1,250 are still represented only by ESTs. Examination of the EST coverage of known cancer-related (CR) genes reveals that <1% do not have corresponding ESTs, indicating that the representation of genes associated with commonly studied tumors is high. The careful recording of the origin of all ESTs we have produced has enabled detailed definition of where the genes they represent are expressed in the human body. More than 100,000 ESTs are available for seven tissues, indicating a surprising variability of gene usage that has led to the discovery of a significant number of genes with restricted expression, and that may thus be therapeutically useful. The ESTs also reveal novel nonsynonymous germline variants (although the one-pass nature of the data necessitates careful validation) and many alternatively spliced transcripts. Although widely exploited by the scientific community, vindicating our totally open source policy, the EST data generated still provide extensive information that remains to be systematically explored, and that may further facilitate progress toward both the understanding and treatment of human cancers.
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http://dx.doi.org/10.1073/pnas.1233632100DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC263829PMC
November 2003

Characterization of COX19, a widely distributed gene required for expression of mitochondrial cytochrome oxidase.

J Biol Chem 2002 Oct 8;277(43):40206-11. Epub 2002 Aug 8.

Instituto de Pesquisa e Desenvolvimento, Universidade do Vale do Paraiba, São José dos Campos, Brazil 12244-000.

COX19, a nuclear gene of Saccharomyces cerevisiae, was cloned by transformation of a respiratory-deficient mutant from complementation group G188 of a pet mutant collection. The gene codes for an 11-kDa protein (Cox19p) required for expression of cytochrome oxidase. Because cox19 mutants are able to synthesize the mitochondrial and nuclear gene products of cytochrome oxidase, Cox19p probably functions post-translationally during assembly of the enzyme. Cox19p is present in the cytoplasm and mitochondria, where it exists as a soluble intermembrane protein. This dual location is similar to what was previously reported for Cox17p, a low molecular weight copper protein thought to be required for maturation of the CuA center of subunit 2 of cytochrome oxidase. The similarity in their subcellular distribution, combined with the presence of four cysteines in Cox19p that align with a subset of the cysteines in Cox17p, suggests that like the latter, Cox19p may function in metal transport to mitochondria.
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http://dx.doi.org/10.1074/jbc.M207348200DOI Listing
October 2002