title: The exchange factor and diacylglycerol receptor RasGRP3 interacts with dynein light chain 1 through its C-terminal domain.
abstract: RasGRP3 is an exchange factor for Ras-like small GTPases that is activated in response to the second messenger diacylglycerol. As with other diacylglycerol receptors, RasGRP3 is redistributed upon diacylglycerol or phorbol ester binding. Several factors are important in determining the pattern of translocation, including the potency of the diacylglycerol analog, the affinity of the receptor for phospholipids, and in some cases, protein-protein interactions. However, little is known about the mechanisms that play a role in RasGRP3 redistribution aside from the nature of the ligand. To discover potential protein binding partners for RasGRP3, we screened a human brain cDNA library using a yeast two-hybrid approach. We identified dynein light chain 1 as a novel RasGRP3-interacting protein. The interaction was confirmed both in vitro and in vivo and required the C-terminal domain encompassing the last 127 amino acids of RasGRP3. A truncated mutant form of RasGRP3 that lacked this C-terminal domain was unable to interact with dynein light chain 1 and displayed a dramatically altered subcellular localization, with a strong reticular distribution and perinuclear and nuclear localization. These findings suggest that dynein light chain 1 represents a novel anchoring protein for RasGRP3 that may regulate subcellular localization of the exchange factor and, as such, may participate in the signaling mediated by diacylglycerol through RasGRP3.
authors.affiliation: Natural Products and Cancer Biology Program, Cancer Research Center of Hawaii, University of Hawaii, Honolulu, Hawaii 96813, USA.
authors.forename: Sara M
authors.initials: SM
authors.lastname: Okamura
authors.forename: Carolyn E
authors.initials: CE
authors.lastname: Oki-Idouchi
authors.forename: Patricia S
authors.initials: PS
authors.lastname: Lorenzo
mesh_terms: D000818:Animals; D001921:Brain / Q000378:metabolism; D016466:CHO Cells; D002352:Carrier Proteins / Q000737:chemistry*; D006224:Cricetinae; D004075:Diglycerides / Q000737:chemistry; D029721:Drosophila Proteins / Q000737:chemistry*; D004398:Dyneins; D015723:Gene Library; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry / Q000502:physiology*; D006801:Humans; D009154:Mutation; D010703:Phorbol Esters / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D015398:Signal Transduction; D020798:Two-Hybrid System Techniques; D020663:ras Guanine Nucleotide Exchange Factors
chemical_list: D002352:Carrier Proteins; D004075:Diglycerides; D029721:Drosophila Proteins; D020662:Guanine Nucleotide Exchange Factors; D010703:Phorbol Esters; C413212:RASGRP3 protein, human; D020663:ras Guanine Nucleotide Exchange Factors; D004398:Dyneins