Individual palmitoyl residues serve distinct roles in H-ras trafficking, microlocalization, and signaling

被引:164
作者
Roy, S
Plowman, S
Rotblat, B
Prior, IA
Muncke, C
Grainger, S
Parton, RG
Henis, YI
Kloog, Y
Hancock, JF
机构
[1] Univ Queensland, Inst Mol Biolsci, Brisbane, Qld 4072, Australia
[2] Univ Queensland, Ctr Microscopy & Microanal, Brisbane, Qld 4072, Australia
[3] Tel Aviv Univ, Dept Neurobiochem, George S Wise Fac Life Sci, IL-69978 Tel Aviv, Israel
[4] Univ Liverpool, Physiol Lab, Liverpool L69 3BX, Merseyside, England
基金
英国惠康基金;
关键词
D O I
10.1128/MCB.25.15.6722-6733.2005
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
H-ras is anchored to the plasma membrane by two palmitoylated cysteine residues, Cys181 and Cys184, operating in concert with a C-terminal S-farnesyl cysteine carboxymethylester. Here we demonstrate that the two palmitates serve distinct biological roles. Monopalmitoylation of Cys181 is required and sufficient for efficient trafficking of H-ras to the plasma membrane, whereas monopallmitoylation of Cys184 does not permit efficient trafficking beyond the Golgi apparatus. However, once at the plasma membrane, monopalmitoylation of Cys184 supports correct GTP-regulated lateral segregation of H-ras between cbolesterol-dependent and cholesterol-independent microdomains. In contrast, monopallmitoylation of Cys181 dramatically reverses H-ras lateral segregation, driving GTP-loaded H-ras into cholesterol-dependent microdomains. Intriguingly, the Cys181 monopalmitoylated H-ras anchor emulates the GTP-regulated microdomain interactions of N-ras. These results identify N-ras as the Ras isoform that normally signals from lipid rafts but also reveal that spacing between palmitate and prenyl groups influences anchor interactions with the lipid bilayer. This concept is further supported by the different plasma membrane affinities of the monopalmitoylated anchors: Cys181-palmitate is equivalent to the dually palmitoylated wild-type anchor, whereas Cys184-pahnitate is weaker. Thus, membrane affinity of a pallmitoylated anchor is a function both of the hydrophobicity of the lipid moieties and their spatial organization. Finally we show that the plasma membrane affinity of monopahnitoylated anchors is absolutely dependent on cholesterol, identifying a new role for cholesterol in promoting interactions with the raft and nonraft plasma membrane.
引用
收藏
页码:6722 / 6733
页数:12
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