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PMID: 18466747 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

A yeast genetic screen reveals a critical role for the pore helix domain in TRP channel gating.

Neuron ·Vol. 58 ·No. 3 ·2008-05-08 ·Pages 362-73

Myers BR, Bohlen CJ, Julius D

Abstract

TRP cation channels function as cellular sensors in uni- and multicellular eukaryotes. Despite intensive study, the mechanisms of TRP channel activation by chemical or physical stimuli remain poorly understood. To identify amino acid residues crucial for TRP channel gating, we developed an unbiased, high-throughput genetic screen in yeast that uncovered rare, constitutively active mutants of the capsaicin receptor, TRPV1. We show that mutations within the pore helix domain dramatically increase basal channel activity and responsiveness to chemical and thermal stimuli. Mutation of corresponding residues within two related TRPV channels leads to comparable effects on their activation properties. Our data suggest that conformational changes in the outer pore region are critical for determining the balance between open and closed states, providing evidence for a general role for this domain in TRP channel activation.

MeSH Terms
Animals Capsaicin/pharmacology Cell Line Genetic Testing/methods Hot Temperature Humans Ion Channel Gating/physiology Kidney/cytology Membrane Potentials/drug effects,physiology Mutagenesis/physiology Oocytes/physiology Patch-Clamp Techniques Protein Structure, Secondary Rats Saccharomyces cerevisiae Stimulation, Chemical Structure-Activity Relationship TRPV Cation Channels/chemistry,genetics,physiology Xenopus laevis
Chemicals
TRPV Cation Channels Trpv1 protein, rat Capsaicin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Myers Benjamin R
Department of Physiology, University of California, San Francisco, 600 16th Street, San Francisco, CA 94143-2140, USA.
Bohlen Christopher J
Julius David
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Article Info
Journal
Neuron
Abbr.
Neuron
ISSN
1097-4199
DOI
10.1016/j.neuron.2008.04.012
Published
2008-05-08
Pages
362-73
Language
English
Region
United States
NLM ID
8809320
PMCID
PMC2422846
Subset
IM
Grants
NINDS NIH HHS · R37 NS047723-16 · United States
NINDS NIH HHS · R37 NS047723-15 · United States
NINDS NIH HHS · R37 NS047723-13 · United States
NINDS NIH HHS · R37 NS047723 · United States
NINDS NIH HHS · R37 NS047723-14 · United States
External Links
PubMed source DOI: 10.1016/j.neuron.2008.04.012 PMC Full Text View journal details
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