Calero M, Whittaker G R, Collins R N
Department of Molecular Medicine, and Microbiology and Immunology, Cornell University, Ithaca, New York 14830, USA.
J Biol Chem. 2001 Apr 13;276(15):12100-12. doi: 10.1074/jbc.M008439200. Epub 2001 Jan 22.
Rab proteins are small GTPases that are essential elements of the protein transport machinery of eukaryotic cells. Each round of membrane transport requires a cycle of Rab protein nucleotide binding and hydrolysis. We have recently characterized a protein, Yip1p, which appears to play a role in Rab-mediated membrane transport in Saccharomyces cerevisiae. In this study, we report the identification of a Yip1p-associated protein, Yop1p. Yop1p is a membrane protein with a hydrophilic region at its N terminus through which it interacts specifically with the cytosolic domain of Yip1p. Yop1p could also be coprecipitated with Rab proteins from total cellular lysates. The TB2 gene is the human homolog of Yop1p (Kinzler, K. W., Nilbert, M. C., Su, L.-K., Vogelstein, B., Bryan, T. M., Levey, D. B., Smith, K. J., Preisinger, A. C., Hedge, P., McKechnie, D., Finniear, R., Markham, A., Groffen, J., Boguski, M. S., Altschul, S. F., Horii, A., Ando, H. M., Y., Miki, Y., Nishisho, I., and Nakamura, Y. (1991) Science 253, 661-665). Our data demonstrate that Yop1p negatively regulates cell growth. Disruption of YOP1 has no apparent effect on cell viability, while overexpression results in cell death, accumulation of internal cell membranes, and a block in membrane traffic. These results suggest that Yop1p acts in conjunction with Yip1p to mediate a common step in membrane traffic.
Rab蛋白是小GTP酶,是真核细胞蛋白质转运机制的关键组成部分。每一轮膜转运都需要Rab蛋白核苷酸结合与水解的循环。我们最近鉴定了一种蛋白质Yip1p,它似乎在酿酒酵母的Rab介导的膜转运中发挥作用。在本研究中,我们报告了Yip1p相关蛋白Yop1p的鉴定。Yop1p是一种膜蛋白,其N端有一个亲水区域,通过该区域它与Yip1p的胞质结构域特异性相互作用。Yop1p也可以从总细胞裂解物中与Rab蛋白共沉淀。TB2基因是Yop1p的人类同源物(金兹勒,K.W.,尼尔伯特,M.C.,苏,L.-K.,沃格尔斯坦,B.,布莱恩,T.M.,利维,D.B.,史密斯,K.J.,普雷辛格,A.C.,赫奇,P.,麦基尔尼,D.,芬尼尔,R.,马克姆,A.,格罗芬,J.,博古斯基,M.S.,阿尔茨舒尔,S.F.,堀井,A.,安藤,H.M.,Y.,三木,Y.,西岛,I.,和中村,Y.(1991年)《科学》253卷,661 - 665页)。我们的数据表明Yop1p对细胞生长起负调控作用。YOP1的破坏对细胞活力没有明显影响,而过表达则导致细胞死亡、内部细胞膜积累以及膜运输受阻。这些结果表明Yop1p与Yip1p共同作用,介导膜运输中的一个共同步骤。