白色念珠菌CaBEM1基因的克隆及其在酿酒酵母丝状生长中的作用

白色念珠菌在不同的生长条件下能发生显著的形态变化 ,这种变化由多种调控因子与信号转导途径所调控。酿酒酵母的G1期细胞周期蛋白Cln1和Cln2参与其形态发生 ,cln1/cln1、cln2 /cln2双缺失株不能形成菌丝。把白色念珠菌基因组文库导入cln1/cln1、cln2 /cln2缺失株 ,筛选能校正菌丝形成缺陷的基因 ,分离得到白色念珠菌中的CaBEM 1基因。从核苷酸序列推导 ,CaBEM1编码一种 6 32个氨基酸的蛋白质 ,氨基酸序列分析表明在其N端有 2个SH3结构域 ,中部有 1个PX结构域 ,C端有 1个PB1结构域 ;CaBem1的氨基酸序列与酿酒酵母的Bem1同源性达 38% ,与裂殖酵母的Scd2同源性达 32 %。在酿酒酵母的缺失株中异源表达CaBEM1,能够部分校正它们在氮源缺乏条件下的菌丝形成缺陷。这种菌丝形成的校正作用绕过MAPK途径和cAMP/PKA途径 ,表明CaBem1在菌丝形成中的作用可能位于这两条信号转导途径的下游

Cloning of Candida albicans CaBEM1 and Its Role in Filamentous Growth of Saccharomyces cerevisiae

The pathogenic fungus Candida albicans has a dimorphic transition in various environmental conditions. Many regulatory factors and several transduction pathways have been identified in controlling filamentous growth. G 1 cyclins Cln1 and Cln2 have been reported as involved in the control of morphogenesis in Saccharomyces cerevisiae. Diploid cln1/cln1 and cln2/cln2 strains completely lost the ability to form pseudohyphae. A C. albicans genomic DNA library was introduced into cln1/cln1 and cln2/cln2 mutants to screen genes which could complement its filamentous growth defect. In this screening a BEM1 homolog, CaBEM1, was identified. The CaBEM1 gene has an ORF of 1 899 bp, encoding a putative protein of 632 amino acids. The CaBem1 protein shares highest homology in amino acids with Bem1(38%) of S.cerevisiae and Scd2(32%) of Schizosaccharomyces pombe . Sequence analysis showed that the CaBem1 contains two N terminal SH3 domains, a PX domain and a C terminal PB1 domain. It is believed these domains are required for binding to proteins involved in polarized growth in S.cerevisiae and S.pombe. Ectopic expression of the CaBEM1 gene in diploid S. cerevisiae suppressed defects in filamentous growth of some mutants under nitrogen starvation conditions. This suppression bypassed MAPK pathway and cAMP/PKA pathway in filamentous growth. These results suggest that the CaBem1 protein may be a downstream component of these two signal transduction pathways of filament formation.