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The β Subunit of the Heterotrimeric G Protein Triggers the Kluyveromyces lactis Pheromone Response Pathway in the Absence of the Subunit
Rocío Navarro-Olmos*,
Laura Kawasaki*,
Lenin Domínguez-Ramírez,
Laura Ongay-Larios*,
Rosario Pérez-Molina*, and
Roberto Coria*
*Departamento de Genética Molecular, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, México D.F. 04510, Mexico and Department of Molecular and Cellular Biology, College of Biological Sciences, University of California at Davis, Davis, CA 95616
Received for publication June 10, 2009.
Revision received November 24, 2009.
Accepted for publication December 2, 2009.
Monitoring Editor: Charles Boone
Abstract:
The Kluyveromyces lactis heterotrimeric G protein is a canonicalGαβ complex; however, in contrast to Saccharomycescerevisiae, where the G subunit is essential for mating, disruptionof the KlG gene yielded cells with almost intact mating capacity.Expression of a nonfarnesylated G, which behaves as a dominant-negativein S. cerevisiae, did not affect mating in wild-type and G cellsof K. lactis. In contrast to the moderate sterility shown bythe single KlGα, the double KlGα KlG mutant displayedfull sterility. A partial sterile phenotype of the KlG mutantwas obtained in conditions where the KlGβ subunit interacteddefectively with the Gα subunit. The addition of a CCAAXmotif to the C-end of KlGβ, partially suppressed the lackof both KlGα and KlG subunits. In cells lacking KlG, theKlGβ subunit cofractionated with KlGα in the plasmamembrane, but in the KlGα KlG strain was located in thecytosol. When the KlGβ-KlGα interaction was affectedin the KlG mutant, most KlGβ fractionated to the cytosol.In contrast to the generic model of G-protein function, theGβ subunit of K. lactis has the capacity to attach to themembrane and to activate mating effectors in absence of theG subunit.