A phosphatidylinositol 3-kinase of Candida albicans: molecular cloning and characterization

作者：Eck R^*; Bruckmann A; Wetzker R; Kunkel W

来源：Yeast, 2000, 16(10): 933-944.

DOI：10.1002/1097-0061(200007)16:10<933::AID-YEA591>3.3.CO;2-3

• phosphatidylinositol 3-kinase
• growth regulated
• Candida albicans

A phosphatidylinositol (PI) 3-kinase gene (CaVPS34) of the human pathogenic yeast Candida albicans was cloned by a PCR-based homology approach. The open reading frame encodes a 1020 amino acid protein with a calculated molecular weight of 118 kDa and a relative isoelectric point of 6.9. It shares 47% sequence identity with Saccharomyces cerevisiae Vps34p. Southern pattern indicated that CaVPS34 is probably present as a single copy gene per haploid genome in C. albicans. We localized the CaVPS34 gene on chromosome 1. Under all conditions tested a major CaVPS34 transcript of approximately 3.5 kb could be detected. CaVPS34 mRNA levels increased during exponential growth up to 12-fold followed by a decline upon entry into stationary phase. The size of a 6x His tag-CaVps34p fusion protein purified from Escherichia call is in agreement with the calculated molecular mass of CaVps34p. It exhibits bl vine PT 3-kinase activity and produces only phosphatidylinositol 3-phosphate. The CaVPS34 gene under the control of its own promoter were not able to complement the temperature-sensitive growth of S. cerevisiae vps34. However, overexpression of CaVPS34 was sufficient to rescue the temperature-sensitive vps34 phenotype, suggesting a functional conservation in C. albicans. The EMBL Accession No. for the sequence reported in this paper is Y09043.

• 出版日期2000-7