An Acylation Cycle Regulates Localization and Activity of Palmitoylated Ras Isoforms

We show that the specific subcellular distribution of H- and Nras guanosine triphosphate–binding proteins is generated by a constitutive de/reacylation cycle that operates on palmitoylated proteins, driving their rapid exchange between the plasma membrane (PM) and the Golgi apparatus. Depalmitoylation redistributes farnesylated Ras in all membranes, followed by repalmitoylation and trapping of Ras at the Golgi, from where it is redirected to the PM via the secretory pathway. This continuous cycle prevents Ras from nonspecific residence on endomembranes, thereby maintaining the specific intracellular compartmentalization. The de/reacylation cycle also initiates Ras activation at the Golgi by transport of PM-localized Ras guanosine triphosphate. Different de/repalmitoylation kinetics account for isoform-specific activation responses to growth factors.

[1]  L. Xue,et al.  A Palmitoylation Switch Mechanism in the Regulation of the Visual Cycle , 2004, Cell.

[2]  J. B. Sajous,et al.  Ras signalling on the endoplasmic reticulum and the Golgi , 2002, Nature Cell Biology.

[3]  A. Álvarez-Barrientos,et al.  Distance-dependent cellular palmitoylation of de-novo-designed sequences and their translocation to plasma membrane subdomains. , 2002, Journal of cell science.

[4]  A. Hall,et al.  Dynamic fatty acylation of p21N‐ras. , 1987, The EMBO journal.

[5]  Herbert Waldmann,et al.  Bioorganic synthesis of lipid-modified proteins for the study of signal transduction , 2000, Nature.

[6]  A. ten Brinke,et al.  Structural requirements for palmitoylation of surfactant protein C precursor. , 2002, The Biochemical journal.

[7]  J. Hancock,et al.  H-Ras Signaling and K-Ras Signaling Are Differentially Dependent on Endocytosis , 2002, Molecular and Cellular Biology.

[8]  J. Silvius.  Mechanisms of Ras Protein Targeting in Mammalian Cells , 2002, The Journal of Membrane Biology.

[9]  David Michaelson,et al.  Differential Localization of Rho Gtpases in Live Cells , 2001, The Journal of cell biology.

[10]  J. Lippincott-Schwartz,et al.  Kinetic Analysis of Secretory Protein Traffic and Characterization of Golgi to Plasma Membrane Transport Intermediates in Living Cells , 1998, The Journal of cell biology.

[11]  M. Resh,et al.  Inhibition of Protein Palmitoylation, Raft Localization, and T Cell Signaling by 2-Bromopalmitate and Polyunsaturated Fatty Acids* , 2000, The Journal of Biological Chemistry.

[12]  George H. Patterson,et al.  A Photoactivatable GFP for Selective Photolabeling of Proteins and Cells , 2002, Science.

[13]  S. Hofmann,et al.  Depalmitoylation of CAAX Motif Proteins , 1995, The Journal of Biological Chemistry.

[14]  O. Bizzozero,et al.  Structural determinants influencing the reaction of cysteine-containing peptides with palmitoyl-coenzyme A and other thioesters. , 2001, Biochimica et biophysica acta.

[15]  P J Verveer,et al.  Quantitative imaging of lateral ErbB1 receptor signal propagation in the plasma membrane. , 2000, Science.

[16]  J. Hancock,et al.  H-ras but Not K-ras Traffics to the Plasma Membrane through the Exocytic Pathway , 2000, Molecular and Cellular Biology.

[17]  E. Nishida,et al.  Sef is a spatial regulator for Ras/MAP kinase signaling. , 2004, Developmental cell.

[18]  Linyi Chen,et al.  The yeast DHHC cysteine-rich domain protein Akr1p is a palmitoyl transferase , 2002, The Journal of cell biology.

[19]  Andre Hoelz,et al.  Structural Evidence for Feedback Activation by Ras·GTP of the Ras-Specific Nucleotide Exchange Factor SOS , 2003, Cell.

[20]  Peter J. Cullen,et al.  Phospholipase Cγ activates Ras on the Golgi apparatus by means of RasGRP1 , 2003, Nature.

[21]  F. L'Heureux,et al.  Fluorimetric evaluation of the affinities of isoprenylated peptides for lipid bilayers. , 1994, Biochemistry.

[22]  T M Jovin,et al.  Imaging the intracellular trafficking and state of the AB5 quaternary structure of cholera toxin. , 1996, The EMBO journal.

[23]  D. Lowy,et al.  New clue to Ras lipid glue , 1989, Nature.

[24]  S. Michaelis,et al.  Mammalian Prenylcysteine Carboxyl Methyltransferase Is in the Endoplasmic Reticulum* , 1998, The Journal of Biological Chemistry.

[25]  B. Kinsella,et al.  Posttranslational modification of Ha-ras p21 by farnesyl versus geranylgeranyl isoprenoids is determined by the COOH-terminal amino acid. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[26]  C. Marshall,et al.  A polybasic domain or palmitoylation is required in addition to the CAAX motif to localize p21 ras to the plasma membrane , 1990, Cell.

[27]  J. Smotrys,et al.  Palmitoylation of intracellular signaling proteins: regulation and function. , 2004, Annual review of biochemistry.

[28]  D. Bredt,et al.  Protein palmitoylation: a regulator of neuronal development and function , 2002, Nature Reviews Neuroscience.

[29]  J. Hancock,et al.  Ras proteins: different signals from different locations , 2003, Nature Reviews Molecular Cell Biology.

[30]  C. Marshall,et al.  All ras proteins are polyisoprenylated but only some are palmitoylated , 1989, Cell.

[31]  A. Pellicer,et al.  Ras Activation in Jurkat T cells following Low-Grade Stimulation of the T-Cell Receptor Is Specific to N-Ras and Occurs Only on the Golgi Apparatus , 2004, Molecular and Cellular Biology.

[32]  J. Silvius,et al.  Doubly-lipid-modified protein sequence motifs exhibit long-lived anchorage to lipid bilayer membranes. , 1995, Biochemistry.

[33]  Roland Eils,et al.  Global Chromosome Positions Are Transmitted through Mitosis in Mammalian Cells , 2003, Cell.

[34]  S. Michaelis,et al.  Endoplasmic reticulum membrane localization of Rce1p and Ste24p, yeast proteases involved in carboxyl-terminal CAAX protein processing and amino-terminal a-factor cleavage. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[35]  A. Ault,et al.  Erf4p and Erf2p Form an Endoplasmic Reticulum-associated Complex Involved in the Plasma Membrane Localization of Yeast Ras Proteins* , 2002, The Journal of Biological Chemistry.

[36]  H. Waldmann,et al.  S-Acylation and plasma membrane targeting of the farnesylated carboxyl-terminal peptide of N-ras in mammalian fibroblasts. , 1997, Biochemistry.

[37]  R. Parton,et al.  Meeting of the apical and basolateral endocytic pathways of the Madin- Darby canine kidney cell in late endosomes , 1989, The Journal of cell biology.

[38]  M. Marsh,et al.  The on-off story of protein palmitoylation. , 2003, Trends in cell biology.

[39]  T. Morimoto,et al.  Endomembrane Trafficking of Ras The CAAX Motif Targets Proteins to the ER and Golgi , 1999, Cell.