Uemura Hiroshi, Watanabe-Yoshida Miyuki, Ishii Nobuya, Shinzato Tomoko, Haw Robin, Aoki Yuko
National Institute of Advanced Industrial Science and Technology, AIST Tsukuba Central 6, Higashi 1-1-1, Tsukuba, Ibaraki, 305-8566, Japan.
Yeast. 2004 Jan 15;21(1):1-10. doi: 10.1002/yea.1048.
To study the function of RAP1, a Candida albicans gene (CaRAP1) that shows sequence similarity to RAP1 of Saccharomyces cerevisiae was isolated by colony hybridization. DNA sequencing predicted an open reading frame of 429 amino acids with an overall identity of 24% to the ScRap1p. The DNA binding domain (DBD) was highly conserved, and EMSA using a GST-CaRap1p fusion protein confirmed its binding ability to the RPG-box of S. cerevisiae ENO1. In contrast, the N-terminus was less conserved and a moderate homology was observed in the BRCT domain. Interestingly, CaRap1p did not contain the C-terminal activation/repression region of ScRap1p.
为研究RAP1的功能，通过菌落杂交分离出白色念珠菌中一个与酿酒酵母RAP1具有序列相似性的基因（CaRAP1）。DNA测序预测其开放阅读框有429个氨基酸，与ScRap1p的总体一致性为24%。DNA结合结构域（DBD）高度保守，使用GST-CaRap1p融合蛋白进行的电泳迁移率变动分析（EMSA）证实了其与酿酒酵母ENO1的RPG框的结合能力。相比之下，N端保守性较低，在BRCT结构域中观察到中等同源性。有趣的是，CaRap1p不包含ScRap1p的C端激活/抑制区域。
