The DH-PH region of the giant protein UNC-89 activates RHO-1 GTPase in Caenorhabditis elegans body wall muscle.
The DH-PH region of the giant protein UNC-89 activates RHO-1 GTPase in Caenorhabditis elegans body wall muscle.
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DOI：
10.1016/j.jmb.2008.08.083
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发表时间：
2008-11-21
影响因子：
5.6
通讯作者：
Benian, Guy M.
中科院分区：
文献类型：
--
作者：
Qadota, Hiroshi;Blangy, Anne;Xiong, Ge;Benian, Guy M.
秀丽隐杆线虫基因unc-89的突变导致肌肉a带的紊乱。unc-89编码一个巨大的多肽（900 kDa），在其n端含有一个DH和一个PH结构域，这是Rho GTPases的鸟嘌呤核苷酸交换因子（GEF）蛋白的特征。为了获得DH-PH区对特定Rho家族小gtpase具有活性的证据，我们利用酵母三杂交系统进行了实验。UNC-89的DH-PH区与rh -1（秀丽隐杆线虫RhoA）具有交换活性，但与CED-10（秀丽隐杆线虫Rac）、MIG-2（秀丽隐杆线虫RhoG）或CDC-42（秀丽隐杆线虫Cdc42）无交换活性。DH结构域也仅对RHO-1具有类似的活性。体外结合实验证明了UNC-89的DH-PH区与秀丽隐杆线虫Rho GTPases的相互作用。在秀丽隐杆线虫成虫中，rho-1基因的部分敲低显示出肌球蛋白粗丝的紊乱模式，与unc-89 （su75）引起的表型相似，unc-89是一种突变等位基因，其中包含DH-PH区域的所有同工型都缺失。综上所述，我们提出了一个模型，即UNC-89的DH-PH区域激活了RHO-1 GTPase，用于秀丽隐杆线虫肌肉细胞中肌球蛋白细丝的组织。
Mutation of the C. elegans gene unc-89 results in disorganization of muscle A-bands. unc-89 encodes a giant polypeptide (900 kDa) containing at its N-terminus, a DH followed by a PH domain, which is characteristic of guanine nucleotide exchange factor (GEF) proteins for Rho GTPases. To obtain evidence that the DH-PH region has activity toward specific Rho family small GTPases, we conducted an experiment using the yeast three hybrid system. The DH-PH region of UNC-89 has exchange activity for RHO-1 (C. elegans RhoA), but not for CED-10 (C. elegans Rac), MIG-2 (C. elegans RhoG), or CDC-42 (C. elegans Cdc42). The DH domain only also has similar activity for RHO-1. An in vitro binding assay demonstrates interaction between the DH-PH region of UNC-89 and each of the C. elegans Rho GTPases. Partial knock down of rho-1 in C. elegans adults showed a pattern of disorganization of myosin thick filaments, similar to the phenotype caused by unc-89 (su75), a mutant allele in which all of the isoforms containing the DH-PH region are missing. Taken together, we propose a model that the DH-PH region of UNC-89 activates RHO-1 GTPase for organization of myosin filaments in C. elegans muscle cells.
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DOI：
10.1083/jcb.132.5.835
发表时间：
1996-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Benian GM;Tinley TL;Tang X;Borodovsky M
通讯作者：
Borodovsky M
影响因子：
2.7
作者：
Ferrara, Tracey M.;Flaherty, Denise B.;Benian, Guy M.
通讯作者：
Benian, Guy M.
影响因子：
64.5
作者：
NONET, ML;GRUNDAHL, K;RAND, JB
通讯作者：
RAND, JB
影响因子：
64.5
作者：
MILLER, DM;ORTIZ, I;EPSTEIN, HF
通讯作者：
EPSTEIN, HF
影响因子：
3.5
作者：
De Toledo, M;Colombo, K;Blangy, A
通讯作者：
Blangy, A