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Cloning and characterization of the SEC18 gene from Candida albicans
Author(s) -
Nieto Almudena,
Sentandreu Rafael,
Agudo Lucas Del Castillo,
Sanz Pascual
Publication year - 1993
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.320090808
Subject(s) - biology , candida albicans , open reading frame , gene , complementation , peptide sequence , saccharomyces cerevisiae , corpus albicans , akt1s1 , microbiology and biotechnology , gene product , mutant , fungal protein , sequence analysis , biochemistry , genetics , hspa2 , gene expression
The SEC18 gene product is required for protein transport at different stages in the Saccharomyces cerevisiae secretory pathway. The homologous SEC18 gene from Candida albicans has been cloned by complementation of a sec18‐1 S. cerevisiae thermosensitive mutant using a C. albicans genomic library in YRp7. Sequence analysis of the gene revealed a 2382‐bp open reading frame which coded for a protein of 88 926 kDa. By an in vitro transcription–translation coupled reaction of the C. albicans SEC18 gene, a protein of approximately 85 kDa was obtained. Hydrophobicity analysis of the protein did not show any predicted signal sequence nor transmembrane anchor domain. These results and the fact that glycosylation was absent in the protein indicated that C. albicans Sec18p did not enter in the secretory pathway. The alignment of the amino acid sequence revealed that the SEC18 gene from C. albicans was homologous to the SEC18 from S. cerevisiae (50% amino acid identity) and to the gene that coded the N‐ethylmaleimide‐sensitive factor (NSF) protein (43% amino acid identity). Moreover, the C. albicans Sec18p also showed the putative ATP binding site present in S. cerevisiae Sec18p and in NSF.