Institute of Development, Aging and Cancer, Tohoku University, Miyagi, Japan.
EMBO J. 2011 Jan 5;30(1):130-44. doi: 10.1038/emboj.2010.276. Epub 2010 Nov 9.
Proper attachment of microtubules to kinetochores is essential for accurate chromosome segregation. Here, we report a novel protein involved in kinetochore-microtubule attachment, chromosome alignment-maintaining phosphoprotein (CAMP) (C13orf8, ZNF828). CAMP is a zinc-finger protein containing three characteristic repeat motifs termed the WK, SPE, and FPE motifs. CAMP localizes to chromosomes and the spindle including kinetochores, and undergoes CDK1-dependent phosphorylation at multiple sites during mitosis. CAMP-depleted cells showed severe chromosome misalignment, which was associated with the poor resistance of K-fibres to the tension exerted upon establishment of sister kinetochore bi-orientation. We found that the FPE region, which is responsible for spindle and kinetochore localization, is essential for proper chromosome alignment. The C-terminal region containing the zinc-finger domains negatively regulates chromosome alignment, and phosphorylation in the FPE region counteracts this regulation. Kinetochore localization of CENP-E and CENP-F was affected by CAMP depletion, and by expressing CAMP mutants that cannot functionally rescue CAMP depletion, placing CENP-E and CENP-F as downstream effectors of CAMP. These data suggest that CAMP is required for maintaining kinetochore-microtubule attachment during bi-orientation.
微管与动粒的正确连接对于准确的染色体分离至关重要。在这里,我们报告了一种新的参与动粒-微管连接的蛋白质,即染色体排列维持磷酸蛋白(CAMP)(C13orf8,ZNF828)。CAMP 是一种含有三个特征重复基序的锌指蛋白,分别称为 WK、SPE 和 FPE 基序。CAMP 定位于染色体和纺锤体,包括动粒,并在有丝分裂期间经历 CDK1 依赖性的多个位点磷酸化。CAMP 耗竭细胞表现出严重的染色体错位,这与 K 纤维对姐妹动粒双定向建立时施加的张力的抵抗力差有关。我们发现,负责纺锤体和动粒定位的 FPE 区域对于正确的染色体排列是必不可少的。包含锌指结构域的 C 端区域负调控染色体排列,而 FPE 区域的磷酸化抵消了这种调控。CAMP 耗竭会影响 CENP-E 和 CENP-F 的动粒定位,并且通过表达不能有效挽救 CAMP 耗竭的 CAMP 突变体,将 CENP-E 和 CENP-F 作为 CAMP 的下游效应物。这些数据表明,CAMP 对于在双定向过程中维持动粒-微管连接是必需的。