Kilmartin John V
MRC Laboratory of Molecular Biology, Cambridge CB2 2QH, UK.
J Cell Biol. 2003 Sep 29;162(7):1211-21. doi: 10.1083/jcb.200307064. Epub 2003 Sep 22.
Centrins are calmodulin-like proteins present in microtubule-organizing centers. The Saccharomyces cerevisiae centrin, Cdc31p, was functionally tagged with a single Z domain of protein A, and used in pull-down experiments to isolate Cdc31p-binding proteins. One of these, Sfi1p, localizes to the half-bridge of the spindle pole body (SPB), where Cdc31p is also localized. Temperature-sensitive mutants in SFI1 show a defect in SPB duplication and genetic interactions with cdc31-1. Sfi1p contains multiple internal repeats that are also present in a Schizosaccharomyces pombe protein, which also localizes to the SPB, and in several human proteins, one of which localizes close to the centriole region. Cdc31p binds directly to individual Sfi1 repeats in a 1:1 ratio, so a single molecule of Sfi1p binds multiple molecules of Cdc31p. The centrosomal human protein containing Sfi1 repeats also binds centrin in the repeat region, showing that this centrin-binding motif is conserved.
中心蛋白是存在于微管组织中心的类钙调蛋白。酿酒酵母的中心蛋白Cdc31p在功能上被标记了蛋白A的单个Z结构域,并用于下拉实验以分离与Cdc31p结合的蛋白。其中一个蛋白Sfi1p定位于纺锤体极体(SPB)的半桥,Cdc31p也定位于此。SFI1中的温度敏感突变体在SPB复制中表现出缺陷,并与cdc31-1存在遗传相互作用。Sfi1p包含多个内部重复序列,这些重复序列也存在于粟酒裂殖酵母的一种蛋白中,该蛋白也定位于SPB,并且存在于几种人类蛋白中,其中一种定位于靠近中心粒区域处。Cdc31p以1:1的比例直接与单个Sfi1重复序列结合,因此单个Sfi1p分子可结合多个Cdc31p分子。含有Sfi1重复序列的中心体人类蛋白在重复区域也结合中心蛋白,表明这种中心蛋白结合基序是保守的。