QUANTITATIVE ANALYSIS OF SPATIAL REGULATION OF LOW MOLECULAR WEIGHT GTPases AND ITS MEDICAL APPLICATION
Ras and Rho families of GTPases have been implicated in the regulation of a wide variety of cellular signaling. The activity of these GTPases is believed to be regulated strictly by upstream signals, which are mediated by a large number of guanine nucleotide exchange factors (GEFs). However, mechanisms underlying subcellular region-specific regulation of Ras and Rho family members remain largely unknown. As a first step to clarify these issues, we attempted to establish a novel method to detect the active GTP-bound form of the Rho family protein Cdc42 in situ. The Cdc42/Racl interactive binding (CRIB) domain of the Cdc42-specific target ACK1 was used as a probe for detecting the activated form of Cdc42. HeLa cells expressing various mutants of Cdc42 were fixed with paraformaldehyde, and then treated with the CRIB domain of ACK1, which was subsequently visualized by immunoftuorescence microscopy. Indeed, only constitulively activated Cdc42 was detected by the ACK1 CRIB domain. This method is applicable to various mouse and human tissues, and has the potential to become a high-through-put drug screening system.
GTP-binding protein Ras family Rho family Cdc42 GEF subcellular localization
Shuji Ueda Tohru Kataoka Takaya Satoh
Division of Molecular Biology, Department of Molecular and Cellular Biology,Kobe University Graduate School of Medicine, 7-5-1 Kusunoki-cho,Chuo-ku, Kobe 650 - 0017, Japan
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
224-226
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）