Tyrosine phosphorylation of RAS by ABL allosterically enhances effector binding

被引:38
作者
Ting, Pamela Y. [1 ]
Johnson, Christian W. [2 ]
Fang, Cong [3 ,4 ]
Cao, Xiaoqing [1 ]
Graeber, Thomas G. [3 ,4 ]
Mattos, Carla [2 ]
Colicelli, John [1 ]
机构
[1] Univ Calif Los Angeles, David Geffen Sch Med, Dept Biol Chem, Mol Biol Inst,Jonsson Comprehens Canc Ctr, Los Angeles, CA 90095 USA
[2] Northeastern Univ, Dept Chem & Chem Biol, Boston, MA 02115 USA
[3] Univ Calif Los Angeles, Dept Mol & Med Pharmacol, Crump Inst Mol Imaging, Metabol & Prote Ctr,Calif Nanosyst Inst, Los Angeles, CA 90095 USA
[4] Univ Calif Los Angeles, Jonsson Comprehens Canc Ctr, Los Angeles, CA 90095 USA
基金
美国国家卫生研究院; 美国国家科学基金会;
关键词
GTPase; signal transduction; RIN1; RAF1; NUCLEOTIDE EXCHANGE FACTOR; K-RAS; GTPASE ACTIVITY; BCR-ABL; RIN1; PROTEIN; ACTIVATION; PROMOTES; KINASES; SWITCH;
D O I
10.1096/fj.15-271510
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
RAS proteins are signal transduction gatekeepers that mediate cell growth, survival, and differentiation through interactions with multiple effector proteins. The RAS effector RAS- and RAB-interacting protein 1 (RIN1) activates its own downstream effectors, the small GTPase RAB5 and the tyrosine kinase Abelson tyrosine-protein kinase (ABL), to modulate endocytosis and cytoskeleton remodeling. To identify ABL substrates downstream of RAS-to-RIN1 signaling, we examined human HEK293T cells over-expressing components of this pathway. Proteomic analysis revealed several novel phosphotyrosine peptides, including Harvey rat sarcoma oncogene (HRAS)-pTyr(137). Here we report that ABL phosphorylates tyrosine 137 of H-, K-, and NRAS. Increased RIN1 levels enhanced HRAS-Tyr(137) phosphorylation by nearly 5-fold, suggesting that RAS-stimulated RIN1 can drive ABL-mediated RAS modification in a feedback circuit. Tyr(137) is well conserved among RAS orthologs and is part of a transprotein H-bond network. Crystal structures of HRAS(Y137F) and HRAS(Y137E) revealed conformation changes radiating from the mutated residue. Although consistent with Tyr(137) participation in allosteric control of HRAS function, the mutations did not alter intrinsic GTP hydrolysis rates in vitro. HRAS-Tyr(137) phosphorylation enhanced HRAS signaling capacity in cells, however, as reflected by a 4-fold increase in the association of phosphorylated HRAS(G12V) with its effector protein RAF proto-oncogene serine/threonine protein kinase 1 (RAF1). These data suggest that RAS phosphorylation at Tyr(137) allosterically alters protein conformation and effector binding, providing a mechanism for effector-initiated modulation of RAS signaling.
引用
收藏
页码:3750 / 3761
页数:12
相关论文
共 61 条
  • [1] PHENIX: a comprehensive Python']Python-based system for macromolecular structure solution
    Adams, Paul D.
    Afonine, Pavel V.
    Bunkoczi, Gabor
    Chen, Vincent B.
    Davis, Ian W.
    Echols, Nathaniel
    Headd, Jeffrey J.
    Hung, Li-Wei
    Kapral, Gary J.
    Grosse-Kunstleve, Ralf W.
    McCoy, Airlie J.
    Moriarty, Nigel W.
    Oeffner, Robert
    Read, Randy J.
    Richardson, David C.
    Richardson, Jane S.
    Terwilliger, Thomas C.
    Zwart, Peter H.
    [J]. ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2010, 66 : 213 - 221
  • [2] Regulation of the oncogenic activity of BCR-ABL by a tightly bound substrate protein RIN1
    Afar, DEH
    Han, LM
    McLaughlin, J
    Wong, S
    Dhaka, A
    Parmar, K
    Rosenberg, N
    Witte, ON
    Colicelli, J
    [J]. IMMUNITY, 1997, 6 (06) : 773 - 782
  • [3] Regulating the regulator: post-translational modification of RAS
    Ahearn, Ian M.
    Haigis, Kevin
    Bar-Sagi, Dafna
    Philips, Mark R.
    [J]. NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2012, 13 (01) : 39 - 51
  • [4] Confirmation of the arginine-finger hypothesis for the GAP-stimulated GTP-hydrolysis reaction of Ras
    Ahmadian, MR
    Stege, P
    Scheffzek, K
    Wittinghofer, A
    [J]. NATURE STRUCTURAL BIOLOGY, 1997, 4 (09) : 686 - 689
  • [5] Balaji Kavitha, 2013, Commun Integr Biol, V6, pe25421, DOI 10.4161/cib.25421
  • [6] RIN1 orchestrates the activation of RAB5 GTPases and ABL tyrosine kinases to determine the fate of EGFR
    Balaji, Kavitha
    Mooser, Chelsea
    Janson, Christine M.
    Bliss, Joanne M.
    Hojjat, Houmehr
    Colicelli, John
    [J]. JOURNAL OF CELL SCIENCE, 2012, 125 (23) : 5887 - 5896
  • [7] A probability-based approach for high-throughput protein phosphorylation analysis and site localization
    Beausoleil, Sean A.
    Villen, Judit
    Gerber, Scott A.
    Rush, John
    Gygi, Steven P.
    [J]. NATURE BIOTECHNOLOGY, 2006, 24 (10) : 1285 - 1292
  • [8] PKC regulates a farnesyl-electrostatic switch on K-Ras that promotes its association with Bcl-XL on mitochondria and induces apoptosis
    Bivona, TG
    Quatela, SE
    Bodemann, BO
    Ahearn, IM
    Soskis, MJ
    Mor, A
    Miura, J
    Wiener, HH
    Wright, L
    Saba, SG
    Yim, D
    Fein, A
    Perez de Castro, I
    Li, C
    Thompson, CB
    Cox, AD
    Philips, MR
    [J]. MOLECULAR CELL, 2006, 21 (04) : 481 - 493
  • [9] Organic solvents order the dynamic switch II in Ras crystals
    Buhrman, G
    de Serrano, V
    Mattos, C
    [J]. STRUCTURE, 2003, 11 (07) : 747 - 751
  • [10] Analysis of Binding Site Hot Spots on the Surface of Ras GTPase
    Buhrman, Greg
    O'Connor, Casey
    Zerbe, Brandon
    Kearney, Bradley M.
    Napoleon, Raeanne
    Kovrigina, Elizaveta A.
    Vajda, Sandor
    Kozakov, Dima
    Kovrigin, Evgenii L.
    Mattos, Carla
    [J]. JOURNAL OF MOLECULAR BIOLOGY, 2011, 413 (04) : 773 - 789