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Title: Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain

Activation of RAF kinase involves the association of its RAS-binding domain (RBD) and cysteine-rich domain (CRD) with membrane-anchored RAS. However, the overall architecture of the RAS/RBD/CRD ternary complex and the orientations of its constituent domains at the membrane remain unclear. Here in this paper, we have combined all-atom and coarse-grained molecular dynamics (MD) simulations with experimental data to construct and validate a model of membrane-anchored CRD, and used this as a basis to explore models of membrane-anchored RAS/RBD/CRD complex. First, simulations of the CRD revealed that it anchors to the membrane via insertion of its two hydrophobic loops, which is consistent with our NMR measurements of CRD bound to nanodiscs. Simulations of the CRD in the context of membrane-anchored RAS/RBD then show how CRD association with either RAS or RBD could play an unexpected role in guiding the membrane orientations of RAS/RBD. This finding has implications for the formation of RAS-RAS dimers, as different membrane orientations of RAS expose distinct putative dimerization interfaces.
Authors:
ORCiD logo [1] ; ORCiD logo [1] ;  [2] ; ORCiD logo [1] ;  [3] ;  [2] ; ORCiD logo [1]
  1. Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
  2. Leidos Biomedical Research, Inc., Frederick, MD (United States). Frederick National Lab. for Cancer Research
  3. Univ. of Wisconsin, Madison, WI (United States). National Magnetic Resource Facility at Madison, Biochemistry Dept.
Publication Date:
Report Number(s):
LA-UR-18-21268
Journal ID: ISSN 2045-2322
Grant/Contract Number:
AC52-06NA25396; AC52-07NA27344; AC02-06CH11357; HHSN261200800001E; P41GM103399
Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Research Org:
Los Alamos National Lab. (LANL), Los Alamos, NM (United States)
Sponsoring Org:
USDOE Laboratory Directed Research and Development (LDRD) Program; National Institutes of Health (NIH); National Science Foundation (NSF)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Biological Science
OSTI Identifier:
1441351

Travers, Timothy, Lopez Bautista, Cesar Augusto, Van, Que, Neale, Christopher Andrew, Tonelli, Marco, Stephen, Andrew, and Gnanakaran, Sandrasegaram. Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain. United States: N. p., Web. doi:10.1038/s41598-018-26832-4.
Travers, Timothy, Lopez Bautista, Cesar Augusto, Van, Que, Neale, Christopher Andrew, Tonelli, Marco, Stephen, Andrew, & Gnanakaran, Sandrasegaram. Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain. United States. doi:10.1038/s41598-018-26832-4.
Travers, Timothy, Lopez Bautista, Cesar Augusto, Van, Que, Neale, Christopher Andrew, Tonelli, Marco, Stephen, Andrew, and Gnanakaran, Sandrasegaram. 2018. "Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain". United States. doi:10.1038/s41598-018-26832-4. https://www.osti.gov/servlets/purl/1441351.
@article{osti_1441351,
title = {Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain},
author = {Travers, Timothy and Lopez Bautista, Cesar Augusto and Van, Que and Neale, Christopher Andrew and Tonelli, Marco and Stephen, Andrew and Gnanakaran, Sandrasegaram},
abstractNote = {Activation of RAF kinase involves the association of its RAS-binding domain (RBD) and cysteine-rich domain (CRD) with membrane-anchored RAS. However, the overall architecture of the RAS/RBD/CRD ternary complex and the orientations of its constituent domains at the membrane remain unclear. Here in this paper, we have combined all-atom and coarse-grained molecular dynamics (MD) simulations with experimental data to construct and validate a model of membrane-anchored CRD, and used this as a basis to explore models of membrane-anchored RAS/RBD/CRD complex. First, simulations of the CRD revealed that it anchors to the membrane via insertion of its two hydrophobic loops, which is consistent with our NMR measurements of CRD bound to nanodiscs. Simulations of the CRD in the context of membrane-anchored RAS/RBD then show how CRD association with either RAS or RBD could play an unexpected role in guiding the membrane orientations of RAS/RBD. This finding has implications for the formation of RAS-RAS dimers, as different membrane orientations of RAS expose distinct putative dimerization interfaces.},
doi = {10.1038/s41598-018-26832-4},
journal = {Scientific Reports},
number = 1,
volume = 8,
place = {United States},
year = {2018},
month = {5}
}