CoChat
Sign Up
Cited by 11Protein Kinase Regulation and GTPase SignalingCellular transport and secretionProtein Structure and Dynamics

The Structure of the RLIP76 RhoGAP-Ral Binding Domain Dyad: Fixed Position of the Domains Leads to Dual Engagement of Small G Proteins at the Membrane

Open access
Rajasekar, Karthik V., Campbell, Louise J., Nietlispach, Daniel, Owen, Darerca, Mott, Helen R.·2013·Structure
DOI 10.1016/j.str.2013.09.007PMC PMC3852207
Open this paper in CoChat — free accountOpen PDF

Abstract

RLIP76 is an effector for Ral small GTPases, which in turn lie downstream of the master regulator Ras. Evidence is growing that Ral and RLIP76 play a role in tumorigenesis, invasion, and metastasis. RLIP76 contains both a RhoGAP domain and a Ral binding domain (GBD) and is, therefore, a node between Ras and Rho family signaling. The structure of the RhoGAP-GBD dyad reveals that the RLIP76 RhoGAP domain adopts a canonical RhoGAP domain structure and that the linker between the two RLIP76 domains is structured, fixing the orientation of the two domains and allowing RLIP76 to interact with Rho-family GTPases and Ral simultaneously. However, the juxtaposed domains do not influence each other functionally, suggesting that the RLIP76-Ral interaction controls cellular localization and that the fixed orientation of the two domains orientates the RhoGAP domain with respect to the membrane, allowing it to be perfectly poised to engage its target G proteins.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.str.2013.09.007

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

No references could be resolved yet.

Research works printed but not linked: 49

  1. Awasthi S. Cheng J.Z. Singhal S.S. Pandya U. Sharma R. Singh S.V. Zimniak P. Awasthi Y.C. Functional reassembly of ATP-dependent xenobiotic transport by the N- and C-terminal domains of RLIP76 and identification of ATP binding sequences Biochemistry 40 2001 4159 4168 11300797
  2. Barrett T. Xiao B. Dodson E.J. Dodson G. Ludbrook S.B. Nurmahomed K. Gamblin S.J. Musacchio A. Smerdon S.J. Eccleston J.F. The structure of the GTPase-activating domain from p50rhoGAP Nature 385 1997 458 461 9009196
  3. Barton G.J. ALSCRIPT: a tool to format multiple sequence alignments Protein Eng. 6 1993 37 40 8433969
  4. Battiste J.L. Wagner G. Utilization of site-directed spin labeling and high-resolution heteronuclear nuclear magnetic resonance for global fold determination of large proteins with limited nuclear Overhauser effect data Biochemistry 39 2000 5355 5365 10820006
  5. Boissel L. Houssin N. Chikh A. Rynditch A. Van Hove L. Moreau J. Recruitment of Cdc42 through the GAP domain of RLIP participates in remodeling of the actin cytoskeleton and is involved in Xenopus gastrulation Dev. Biol. 312 2007 331 343 17950267
  6. Brünger A.T. Adams P.D. Clore G.M. DeLano W.L. Gros P. Grosse-Kunstleve R.W. Jiang J.S. Kuszewski J. Nilges M. Pannu N.S. Crystallography & NMR System: a new software suite for macromolecular structure determination Acta Crystallogr. D Biol. Crystallogr. 54 1998 905 921 9757107
  7. Cantor S.B. Urano T. Feig L.A. Identification and characterization of Ral-binding protein 1, a potential downstream target of Ral GTPases Mol. Cell. Biol. 15 1995 4578 4584 7623849 PMC230698
  8. Chien Y. White M.A. RAL GTPases are linchpin modulators of human tumour-cell proliferation and survival EMBO Rep. 4 2003 800 806 12856001 PMC1326339
  9. Clore G.M. Gronenborn A.M. Bax A. A robust method for determining the magnitude of the fully asymmetric alignment tensor of oriented macromolecules in the absence of structural information J. Magn. Reson. 133 1998 216 221 9654491
  10. Coon B.G. Burgner J. Camonis J.H. Aguilar R.C. The epsin family of endocytic adaptors promotes fibrosarcoma migration and invasion J. Biol. Chem. 285 2010 33073 33081 20709745 PMC2963371
  11. Di Tommaso P. Moretti S. Xenarios I. Orobitg M. Montanyola A. Chang J.M. Taly J.F. Notredame C. T-Coffee: a web server for the multiple sequence alignment of protein and RNA sequences using structural information and homology extension Nucleic Acids Res. 39 2011 W13 W17 21558174 PMC3125728
  12. Elliot-Smith A.E. Owen D. Mott H.R. Lowe P.N. Double mutant cycle thermodynamic analysis of the hydrophobic Cdc42-ACK protein-protein interaction Biochemistry 46 2007 14087 14099 17999470
  13. Fenwick R.B. Prasannan S. Campbell L.J. Nietlispach D. Evetts K.A. Camonis J. Mott H.R. Owen D. Solution structure and dynamics of the small GTPase RalB in its active conformation: significance for effector protein binding Biochemistry 48 2009 2192 2206 19166349
  14. Fenwick R.B. Campbell L.J. Rajasekar K. Prasannan S. Nietlispach D. Camonis J. Owen D. Mott H.R. The RalB-RLIP76 complex reveals a novel mode of Ral-effector interaction Structure 18 2010 985 995 20696399 PMC4214634
  15. Fidyk N.J. Cerione R.A. Understanding the catalytic mechanism of GTPase-activating proteins: demonstration of the importance of switch domain stabilization in the stimulation of GTP hydrolysis Biochemistry 41 2002 15644 15653 12501193
  16. Fitzkee N.C. Bax A. Facile measurement of 1H-15N residual dipolar couplings in larger perdeuterated proteins J. Biomol. NMR 48 2010 65 70 20694505 PMC2950907
  17. Gildea J.J. Harding M.A. Seraj M.J. Gulding K.M. Theodorescu D. The role of Ral A in epidermal growth factor receptor-regulated cell motility Cancer Res. 62 2002 982 985 11861368
  18. Hamad N.M. Elconin J.H. Karnoub A.E. Bai W. Rich J.N. Abraham R.T. Der C.J. Counter C.M. Distinct requirements for Ras oncogenesis in human versus mouse cells Genes Dev. 16 2002 2045 2057 12183360 PMC186434
  19. Houben K. Dominguez C. van Schaik F.M.A. Timmers H.T.M. Bonvin A.M.J.J. Boelens R. Solution structure of the ubiquitin-conjugating enzyme UbcH5B J. Mol. Biol. 344 2004 513 526 15522302
  20. Ikeda M. Ishida O. Hinoi T. Kishida S. Kikuchi A. Identification and characterization of a novel protein interacting with Ral-binding protein 1, a putative effector protein of Ral J. Biol. Chem. 273 1998 814 821 9422736
  21. Jelen F. Lachowicz P. Apostoluk W. Mateja A. Derewenda Z.S. Otlewski J. Dissecting the thermodynamics of GAP-RhoA interactions J. Struct. Biol. 165 2009 10 18 18929667 PMC2656442
  22. Jullien-Flores V. Dorseuil O. Romero F. Letourneur F. Saragosti S. Berger R. Tavitian A. Gacon G. Camonis J.H. Bridging Ral GTPase to Rho pathways. RLIP76, a Ral effector with CDC42/Rac GTPase-activating protein activity J. Biol. Chem. 270 1995 22473 22477 7673236
  23. Jullien-Flores V. Mahé Y. Mirey G. Leprince C. Meunier-Bisceuil B. Sorkin A. Camonis J.H. RLIP76, an effector of the GTPase Ral, interacts with the AP2 complex: involvement of the Ral pathway in receptor endocytosis J. Cell Sci. 113 2000 2837 2844 10910768
  24. Kashatus D.F. Lim K.H. Brady D.C. Pershing N.L.K. Cox A.D. Counter C.M. RALA and RALBP1 regulate mitochondrial fission at mitosis Nat. Cell Biol. 13 2011 1108 1115 21822277 PMC3167028
  25. Lim K.H. Brady D.C. Kashatus D.F. Ancrile B.B. Der C.J. Cox A.D. Counter C.M. Aurora-A phosphorylates, activates, and relocalizes the small GTPase RalA Mol. Cell. Biol. 30 2010 508 523 19901077 PMC2798468
  26. Linge J.P. Habeck M. Rieping W. Nilges M. ARIA: automated NOE assignment and NMR structure calculation Bioinformatics 19 2003 315 316 12538267
  27. Nassar N. Hoffman G.R. Manor D. Clardy J.C. Cerione R.A. Structures of Cdc42 bound to the active and catalytically compromised forms of Cdc42GAP Nat. Struct. Biol. 5 1998 1047 1052 9846874
  28. Neel N.F. Rossman K.L. Martin T.D. Hayes T.K. Yeh J.J. Der C.J. The RalB small GTPase mediates formation of invadopodia through a GTPase-activating protein-independent function of the RalBP1/RLIP76 effector Mol. Cell. Biol. 32 2012 1374 1386 22331470 PMC3318593
  29. Owen D. Mott H.R. Laue E.D. Lowe P.N. Residues in Cdc42 that specify binding to individual CRIB effector proteins Biochemistry 39 2000 1243 1250 10684602
  30. Owen D. Lowe P.N. Nietlispach D. Brosnan C.E. Chirgadze D.Y. Parker P.J. Blundell T.L. Mott H.R. Molecular dissection of the interaction between the small G proteins Rac1 and RhoA and protein kinase C-related kinase 1 (PRK1) J. Biol. Chem. 278 2003 50578 50587 14514689
  31. Owen D. Campbell L.J. Littlefield K. Evetts K.A. Li Z.G. Sacks D.B. Lowe P.N. Mott H.R. The IQGAP1-Rac1 and IQGAP1-Cdc42 interactions: interfaces differ between the complexes J. Biol. Chem. 283 2008 1692 1704 17984089
  32. Park S.H. Weinberg R.A. A putative effector of Ral has homology to Rho/Rac GTPase activating proteins Oncogene 11 1995 2349 2355 8570186
  33. Pirruccello M. Swan L.E. Folta-Stogniew E. De Camilli P. Recognition of the F&H motif by the Lowe syndrome protein OCRL Nat. Struct. Mol. Biol. 18 2011 789 795 21666675 PMC3130824
  34. Rajasekar K.V. Campbell L.J. Nietlispach D. Owen D. Mott H.R. 1H, 13C and 15N resonance assignments of the GTPase-activating (GAP) and Ral binding domains (GBD) of RLIP76 (RalBP1) Biomol. NMR Assign. 6 2012 119 122 21915608
  35. Rittinger K. Walker P.A. Eccleston J.F. Nurmahomed K. Owen D. Laue E. Gamblin S.J. Smerdon S.J. Crystal structure of a small G protein in complex with the GTPase-activating protein rhoGAP Nature 388 1997 693 697 9262406
  36. Rossé C. L’Hoste S. Offner N. Picard A. Camonis J. RLIP, an effector of the Ral GTPases, is a platform for Cdk1 to phosphorylate Epsin during the switch off of endocytosis in mitosis J. Biol. Chem. 278 2003 30597 30604 12775724
  37. Shen Y. Delaglio F. Cornilescu G. Bax A. TALOS+: a hybrid method for predicting protein backbone torsion angles from NMR chemical shifts J. Biomol. NMR 44 2009 213 223 19548092 PMC2726990
  38. Smith S.C. Oxford G. Baras A.S. Owens C. Havaleshko D. Brautigan D.L. Safo M.K. Theodorescu D. Expression of Ral GTPases, their effectors, and activators in human bladder cancer Clin. Cancer Res. 13 2007 3803 3813 17606711
  39. Thompson G. Owen D. Chalk P.A. Lowe P.N. Delineation of the Cdc42/Rac-binding domain of p21-activated kinase Biochemistry 37 1998 7885 7891 9601050
  40. Vatsyayan R. Lelsani P.C.R. Awasthi S. Singhal S.S. RLIP76: a versatile transporter and an emerging target for cancer therapy Biochem. Pharmacol. 79 2010 1699 1705 20097178 PMC2874070
  41. Vranken W.F. Boucher W. Stevens T.J. Fogh R.H. Pajon A. Llinas M. Ulrich E.L. Markley J.L. Ionides J. Laue E.D. The CCPN data model for NMR spectroscopy: development of a software pipeline Proteins 59 2005 687 696 15815974
  42. Wang Z.X. An exact mathematical expression for describing competitive binding of two different ligands to a protein molecule FEBS Lett. 360 1995 111 114 7875313
  43. Ward Y. Wang W. Woodhouse E. Linnoila I. Liotta L. Kelly K. Signal pathways which promote invasion and metastasis: critical and distinct contributions of extracellular signal-regulated kinase and Ral-specific guanine exchange factor pathways Mol. Cell. Biol. 21 2001 5958 5969 11486034 PMC87314
  44. Winn M.D. Ballard C.C. Cowtan K.D. Dodson E.J. Emsley P. Evans P.R. Keegan R.M. Krissinel E.B. Leslie A.G. McCoy A. Overview of the CCP4 suite and current developments Acta Crystallogr. D Biol Crystallogr. 67 2011 235 242 21460441 PMC3069738
  45. Wu Z. Owens C. Chandra N. Popovic K. Conaway M. Theodorescu D. RalBP1 is necessary for metastasis of human cancer cell lines Neoplasia 12 2010 1003 1012 21170262 PMC3003135
  46. Yamaguchi A. Urano T. Goi T. Feig L.A. An Eps homology (EH) domain protein that binds to the Ral-GTPase target, RalBP1 J. Biol. Chem. 272 1997 31230 31234 9395447
  47. Zhang B. Wang Z.X. Zheng Y. Characterization of the interactions between the small GTPase Cdc42 and its GTPase-activating proteins and putative effectors. Comparison of kinetic properties of Cdc42 binding to the Cdc42-interactive domains J. Biol. Chem. 272 1997 21999 22007 9268338
  48. Zhang B. Chernoff J. Zheng Y. Interaction of Rac1 with GTPase-activating proteins and putative effectors. A comparison with Cdc42 and RhoA J. Biol. Chem. 273 1998 8776 8782 9535855
  49. Zweckstetter M. NMR: prediction of molecular alignment from structure using the PALES software Nat. Protoc. 3 2008 679 690 18388951

Cited by

Sorted by citations

Authors
Rajasekar, K. V., Campbell, L. J., Nietlispach, D., Owen, D., & Mott, H. R.
Affiliations
University of Cambridge (GB)
Corresponding author
  • Owen, Darerca
  • Mott, Helen R.
ORCID
Venue
Structure
Publisher
Elsevier BV
Published
2013-12
Volume
21
Issue
12
Pages
2131-2142
ISSN
0969-2126 (ISSN-L)
DOI
10.1016/j.str.2013.09.007
PMC
PMC3852207
OpenAlex
W2026557548
Cited by
11
References
0
Citations by year
2026: 1 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 2 · 2018: 1 · 2017: 2 · 2016: 1 · 2014: 1
FWCI
0.25
Type
journal article
Language
en
Open access
Licence
Locations
  • Structure (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • Wellcome Trust
  • Medical Research Council (MR/J007803/1)
  • Biotechnology and Biological Sciences Research Council (BB/E013228/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
Keywords
RLIP76, Ras signaling, Rho GTPases, membrane targeting, tumor invasion and metastasis, protein structure
MeSH terms
Amino Acid Sequence, Cell Membrane, Humans, Models, Molecular, Molecular Sequence Data, Protein Binding, Protein Structure, Tertiary, ATP-Binding Cassette Transporters, ras Proteins, GTP-Binding Proteins, GTPase-Activating Proteins, ral GTP-Binding Proteins, cdc42 GTP-Binding Protein, rac1 GTP-Binding Protein

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review