Biochemical and molecular studies of the mitochondrial components of Aspergillus fumigatus: Cloning and heterologous expression of NAD/NADH carrier.

• Grant number: 12/15861-9
• Support Opportunities: Scholarships in Brazil - Master
• Effective date (Start): November 01, 2012
• Effective date (End): June 30, 2014
• Field of knowledge: Biological Sciences - Biochemistry - Metabolism and Bioenergetics
• Principal Investigator: Sergio Akira Uyemura
• Grantee: Laís de Lourdes de Lima Bálico
• Host Institution: Faculdade de Ciências Farmacêuticas de Ribeirão Preto (FCFRP). Universidade de São Paulo (USP). Ribeirão Preto , SP, Brazil

Abstract

In the last ten years, our laboratory has been studying the mitochondrial bioenergetics of Aspergillus fumigatus. We have shown the presence of classical components of the respiratory chain (or via of cytochrome c) in this microorganism. In addition, this fungus presents alternative components as alternative NADH dehydrogenase, an alternative oxidase and uncoupling protein-like. In addition to the input of NADH to the respiratory chain via classical and alternative NADH dehydrogenase, studies have identified and characterized two isoforms of mitochondrial protein carrier NAD+ in Arabidopsis thaliana (Ndt1p and Ndt2p, respectively). In previous studies in our laboratory, we observed that in mitochondria of the Paracoccidioides brasiliensis, the mitochondrial membrane potential could be induced by NAD+ and dissipated by uncoupler FCCP, suggesting the presence of a carrier NAD+, as described in S. cerevisiae. Other experiments were performed with mitochondria from A. fumigatus and also suggested the presence of this carrier in this organism. Using bioinformatic approaches, we found a sequence in the genome of A. fumigatus, which has 32% identity with the nicotinamide dinucleotide transporter gene (ndt1p) of S. cerevisiae. The objective of this project is cloning, sequence and heterologous expression of the ndt1p-like gene from A. fumigatus in the S.cerevisiae mutant strain (deltandt1deltandt2).

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DE LIMA BALICO, LAIS DE LOURDES; SANTOS, EMERSON DE SOUZA; SUZUKI-HATANO, SILVELI; SOUSA, LUCAS OLIVEIRA; CALEIRO SEIXAS AZZOLINI, ANA ELISA; LUCISANO-VALIM, YARA MARIA; DINAMARCO, TAISA MAGNANI; KANNEN, VINICIUS; UYEMURA, SERGIO AKIRA. Heterologous expression of mitochondrial nicotinamide adenine dinucleotide transporter (Ndt1) from Aspergillus fumigatus rescues impaired growth in Delta ndt1 Delta ndt2 Saccharomyces cerevisiae strain. Journal of Bioenergetics and Biomembranes, v. 49, n. 6, p. 423-435, DEC 2017. (12/15861-9, 13/20031-8)

Academic Publications

(References retrieved automatically from State of São Paulo Research Institutions)

BÁLICO, Laís de Lourdes de Lima. Molecular and biochemical characterization of a mitochondrial nicotinamide adenine dinucleotide carrier of Aspergillus fumigatus. 2014. Master's Dissertation - Universidade de São Paulo (USP). Faculdade de Ciências Farmacêuticas de Ribeirão Preto (PCARP/BC) Ribeirão Preto.