Differential effects of prenylation and s-acylation on type I and II ROPS membrane interaction and function.
Prenylation primarily by geranylgeranylation is required for membrane attachment and function of type I Rho of Plants (ROPs) and Gγ proteins, while type II ROPs are attached to the plasma membrane by S-acylation. Yet, it is not known how prenylation affects ROP membrane interaction dynamics and what are the functional redundancy and specificity of type I and type II ROPs. Here, we have used the expression of ROPs in mammalian cells together with geranylgeranylation and CaaX prenylation-deficient mutants to answer these questions. Our results show that the mechanism of type II ROP S-acylation and membrane attachment is unique to plants and likely responsible for the viability of plants in the absence of CaaX prenylation activity. The prenylation of ROPs determines their steady-state distribution between the plasma membrane and the cytosol but has little effect on membrane interaction dynamics. In addition, the prenyl group type has only minor effects on ROP function. Phenotypic analysis of the CaaX prenylation-deficient pluripetala mutant epidermal cells revealed that type I ROPs affect cell structure primarily on the adaxial side, while type II ROPs are functional and induce a novel cell division phenotype in this genetic background. Taken together, our studies show how prenyl and S-acyl lipid modifications affect ROP subcellular distribution, membrane interaction dynamics, and function.
Department of Molecular Biology and Ecology of Plants , George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv 69978, Israel.
Nadav
N
Sorek
Orit
O
Gutman
Einat
E
Bar
Mohamad
M
Abu-Abied
Xuehui
X
Feng
Mark P
MP
Running
Efraim
E
Lewinsohn
Naomi
N
Ori
Einat
E
Sadot
Yoav I
YI
Henis
Shaul
S
Yalovsky
D000215:Acylation; D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry*; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D008401:Gas Chromatography-Mass Spectrometry; D007313:Insecta / Q000166:cytology; D008565:Membrane Proteins / Q000737:chemistry*; D051379:Mice; D020559:Monomeric GTP-Binding Proteins / Q000737:chemistry*; D009154:Mutation; D041681:NIH 3T3 Cells; D010641:Phenotype; D019441:Plant Epidermis / Q000166:cytology; D017368:Protein Prenylation
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; C518506:ROP6 protein, Arabidopsis; D020559:Monomeric GTP-Binding Proteins