한빛사논문
Abstract
Donha Parka,1,2, Inish O'Dohertyb,c,3, Rishi K. Somvanshid,3, Axel Bethkeb,c, Frank C. Schroederb,c,2, Ujendra Kumard, and Donald L. Riddlea
aMichael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada, V6T 1Z4;
bBoyce Thompson Institute and
cDepartment of Chemistry and Chemical Biology, Cornell University, Ithaca, NY 14853; and
dFaculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, BC, Canada V6T1Z3
Abstract
A chemically diverse family of small-molecule signals, the ascarosides, control developmental diapause (dauer), olfactory learning, and social behaviors of the nematode model organism, Caenorhabditis elegans. The ascarosides act upstream of conserved signaling pathways, including the insulin, TGF-β, serotonin, and guanylyl cyclase pathways; however, the sensory processes underlying ascaroside function are poorly understood. Because ascarosides often are multifunctional and show strongly synergistic effects, characterization of their receptors will be essential for understanding ascaroside biology and may provide insight into molecular mechanisms that produce synergistic outcomes in small-molecule sensing. Based on DAF-8 immunoprecipitation, we here identify two G-protein-coupled receptors, DAF-37 and DAF-38, which cooperatively mediate ascaroside perception. daf-37 mutants are defective in all responses to ascr#2, one of the most potent dauer-inducing ascarosides, although this mutant responds normally to other ascarosides. In contrast, daf-38 mutants are partially defective in responses to several different ascarosides. Through cell-specific overexpression, we show that DAF-37 regulates dauer when expressed in ASI neurons and adult behavior when expressed in ASK neurons. Using a photoaffinity-labeled ascr#2 probe and amplified luminescence assays (AlphaScreen), we demonstrate that ascr#2 binds to DAF-37. Photobleaching fluorescent energy transfer assays revealed that DAF-37 and DAF-38 form heterodimers, and we show that heterodimerization strongly increases cAMP inhibition in response to ascr#2. These results suggest that that the ascarosides' intricate signaling properties result in part from the interaction of highly structure-specific G-protein?coupled receptors such as DAF-37 with more promiscuous G-protein?coupled receptors such as DAF-38.
chemosensation, photoaffinity labeling, signal transduction, small-molecule receptor, pheromone signaling
Footnotes
1Present address: Centre for Molecular Medicine and Therapeutics, Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada V5Z 4H4.
2To whom correspondence may be addressed.
3I.O. and R.K.S. contributed equally to this work.
논문정보
관련 링크
연구자 키워드
연구자 ID
관련분야 연구자보기
소속기관 논문보기
관련분야 논문보기
해당논문 저자보기