Characterization of a gene which encodes a mannosyltransferase homolog of Paracoccidioides brasiliensis

We screened an expression library of the yeast form of Paracoccidioides brasiliensis with a pool of human sera that was pre-adsorbed with mycelium, from patients with paracoccidioidomycosis (PCM). A sequence ( PbYmnt) was obtained and characterized. A genomic clone was obtained by PCR of P.  brasili...

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Bibliographic Details
Published in:Microbes and infection 2002-08, Vol.4 (10), p.1027-1034
Main Authors: Costa, Alessandra A., Gómez, Francisco J., Pereira, Maristela, Felipe, M.Sueli S., Jesuino, Rosália S.A., Deepe, George S., de Almeida Soares, Célia M.
Format: Article
Language:English
Subjects:
Amino Acid Motifs
Amino Acid Sequence
Antigens
Base Sequence
Biological and medical sciences
Cloning, Molecular
Fundamental and applied biological sciences. Psychology
Fungal Proteins - chemistry
Fungal Proteins - genetics
Fungal Proteins - immunology
Fungal Proteins - isolation & purification
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Humans
Hydrophobic and Hydrophilic Interactions
Mannosyltransferases
Mannosyltransferases - chemistry
Mannosyltransferases - genetics
Mannosyltransferases - immunology
Mannosyltransferases - isolation & purification
Microbiology
Molecular Sequence Data
Mycology
Paracoccidioides - enzymology
Paracoccidioides - genetics
Paracoccidioides - immunology
Paracoccidioides brasiliensis
Paracoccidioidomycosis - immunology
Polymerase Chain Reaction
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Sequence Analysis, DNA
Sequence Homology, Amino Acid
Tropical medicine
Yeast phase
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Summary:We screened an expression library of the yeast form of Paracoccidioides brasiliensis with a pool of human sera that was pre-adsorbed with mycelium, from patients with paracoccidioidomycosis (PCM). A sequence ( PbYmnt) was obtained and characterized. A genomic clone was obtained by PCR of P.  brasiliensis total DNA. The sequence contained a single open reading frame (ORF) encoding a protein of 357 amino acid residues, with a molecular mass of 39.78 kDa. The deduced amino acid sequence exhibited identity to mannosyl- and glycosyltransferases from several sources. A DXD motif was present in the translated gene and this sequence is characteristic of the glycosyltransferases. Hydropathy analysis revealed a single transmembrane region near the amino terminus of the molecule that suggested a type II membrane protein. The PbYmnt was expressed preferentially in the yeast parasitic phase. The accession number of the nucleotide sequence of PbYmnt and its flanking regions is AF374353. A recombinant protein was generated in Escherichia coli. Our data suggest that PbYmnt encodes one member of a glycosyltransferase family of proteins and that our strategy was useful in the isolation of differentially expressed genes.
ISSN:1286-4579
1769-714X
DOI:10.1016/S1286-4579(02)01626-X