Raymond M, Dignard D, Alarco A M, Clark K L, Weber S, Whiteway M, Leberer E, Thomas D Y
Institut de recherches cliniques de Montréal, Montréal, Québec, Canada.
Yeast. 2000 Apr;16(6):531-8. doi: 10.1002/(SICI)1097-0061(200004)16:6<531::AID-YEA543>3.0.CO;2-J.
In a screen for Candida albicans genes capable of supressing a ste20Delta mutation in Saccharomyces cerevisiae, a homologue of the exportin-encoding gene CRM1 was isolated. The CaCRM1 gene codes for a protein of 1079 amino acids with a predicted molecular weight of 124 029 and isoelectric point of 5.04. Crm1p from C. albicans displays significant amino acid sequence homology with Crm1p from Saccharomyces cerevisiae (65% identity, 74% similarity), Schizosaccharomyces pombe (55% identity, 66% similarity), Caenorhabditis elegans (45% identity, 57% similarity), and Homo sapiens (48% identity, 59% similarity). Interestingly, CaCRM1 encodes a threonine rather than a cysteine at position 533 in the conserved central region, suggesting that CaCrm1p is leptomycin B-insensitive, like S. cerevisiae Crm1p. CaCRM1 on a high copy vector can complement a thermosensitive allele of CRM1 (xpo1-1) in S. cerevisiae, showing that CaCrm1p and S. cerevisiae Crm1p are functionally conserved. Southern blot analysis suggests that CaCRM1 is present at a single locus within the C. albicans genome. The nucleotide sequence of the CaCRM1 gene has been deposited at GenBank under Accession No. AF178855.
在一项筛选能够抑制酿酒酵母中ste20Delta突变的白色念珠菌基因的研究中,分离出了输出蛋白编码基因CRM1的一个同源物。CaCRM1基因编码一种由1079个氨基酸组成的蛋白质,预测分子量为124029,等电点为5.04。白色念珠菌的Crm1p与酿酒酵母的Crm1p(65%同一性,74%相似性)、粟酒裂殖酵母的Crm1p(55%同一性,66%相似性)、秀丽隐杆线虫的Crm1p(45%同一性,57%相似性)和智人的Crm1p(48%同一性,59%相似性)显示出显著的氨基酸序列同源性。有趣的是,CaCRM1在保守的中央区域第533位编码的是苏氨酸而非半胱氨酸,这表明CaCrm1p像酿酒酵母的Crm1p一样对雷帕霉素B不敏感。高拷贝载体上的CaCRM1可以互补酿酒酵母中CRM1的一个温度敏感等位基因(xpo1-1),这表明CaCrm1p和酿酒酵母的Crm1p在功能上是保守的。Southern印迹分析表明CaCRM1存在于白色念珠菌基因组中的一个单一基因座上。CaCRM1基因的核苷酸序列已提交至GenBank,登录号为AF178855。