title: Structural analysis of autoinhibition in the Ras-specific exchange factor RasGRP1.
abstract: RasGRP1 and SOS are Ras-specific nucleotide exchange factors that have distinct roles in lymphocyte development. RasGRP1 is important in some cancers and autoimmune diseases but, in contrast to SOS, its regulatory mechanisms are poorly understood. Activating signals lead to the membrane recruitment of RasGRP1 and Ras engagement, but it is unclear how interactions between RasGRP1 and Ras are suppressed in the absence of such signals. We present a crystal structure of a fragment of RasGRP1 in which the Ras-binding site is blocked by an interdomain linker and the membrane-interaction surface of RasGRP1 is hidden within a dimerization interface that may be stabilized by the C-terminal oligomerization domain. NMR data demonstrate that calcium binding to the regulatory module generates substantial conformational changes that are incompatible with the inactive assembly. These features allow RasGRP1 to be maintained in an inactive state that is poised for activation by calcium and membrane-localization signals. DOI:http://dx.doi.org/10.7554/eLife.00813.001.
authors.affiliation: Department of Molecular and Cell Biology , University of California, Berkeley , Berkeley , United States ; California Institute for Quantitative Biosciences , University of California, Berkeley , Berkeley , United States.
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Iwig
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Vercoulen
authors.forename: Rahul
authors.initials: R
authors.lastname: Das
authors.forename: Tiago
authors.initials: T
authors.lastname: Barros
authors.forename: Andre
authors.initials: A
authors.lastname: Limnander
authors.forename: Yan
authors.initials: Y
authors.lastname: Che
authors.forename: Jeffrey G
authors.initials: JG
authors.lastname: Pelton
authors.forename: David E
authors.initials: DE
authors.lastname: Wemmer
authors.forename: Jeroen P
authors.initials: JP
authors.lastname: Roose
authors.forename: John
authors.initials: J
authors.lastname: Kuriyan
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D004268:DNA-Binding Proteins; D020662:Guanine Nucleotide Exchange Factors; C491790:RASGRP1 protein, human; D011994:Recombinant Proteins; D002118:Calcium
keywords: Human; cell signaling; diacylglycerol; nucleotide exchange factor
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