Leung T, How B E, Manser E, Lim L
Institute of Molecular and Cell Biology, National University of Singapore.
J Biol Chem. 1993 Feb 25;268(6):3813-6.
Diverse GTPase-activating proteins (GAPs) for the p21rho subfamily were detected by a novel overlay assay (Manser, E., Leung, T., Monfries, C., Teo, M., Hall, C., and Lim, L. (1992) J. Biol. Chem. 267, 16025-16028), with some GAPs being tissue-specific. Using a PCR strategy exploiting conserved regions common to rho/rac-GAPs, we have isolated a rat testis cDNA encoding a 34-kDa rac-GAP termed beta-chimaerin, as it was highly related to n-chimaerin, containing both a GAP domain (77% identity) and the phorbol ester-binding region (93% identity). beta-Chimaerin mRNA is expressed exclusively in the testis at the onset of sexual maturation. In situ hybridization and cell fractionation analyses show beta-chimaerin mRNA expression to be stage-specific, paralleling acrosomal assembly at the late stage of spermatogenesis. A corresponding testis-specific 30-kDa rac-GAP was detected. The testis-specific and stage-dependent expression of this new member of the chimaerin family offer an alternative model system for investigating the functional role of this class of p21 GAPs, particularly in relation to cytoskeletal reorganization.
通过一种新型覆盖分析方法(曼泽尔,E.,梁,T.,蒙弗里斯,C.,特奥,M.,霍尔,C.,和林,L.(1992年)《生物化学杂志》267卷,16025 - 16028页)检测到了针对p21rho亚家族的多种GTP酶激活蛋白(GAPs),其中一些GAPs具有组织特异性。利用一种PCR策略,该策略利用了rho/rac - GAPs共有的保守区域,我们分离出了一个大鼠睾丸cDNA,它编码一种34 kDa的rac - GAP,称为β - 嵌合蛋白,因为它与n - 嵌合蛋白高度相关,同时包含一个GAP结构域(77%的同一性)和佛波酯结合区域(93%的同一性)。β - 嵌合蛋白mRNA在性成熟开始时仅在睾丸中表达。原位杂交和细胞分级分离分析表明,β - 嵌合蛋白mRNA的表达具有阶段特异性,与精子发生后期的顶体组装平行。还检测到了一种相应的睾丸特异性30 kDa的rac - GAP。嵌合蛋白家族这一新成员的睾丸特异性和阶段依赖性表达为研究这类p21 GAPs的功能作用提供了一个替代模型系统,特别是与细胞骨架重组相关的功能作用。