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Systematic analysis of essential genes reveals new regulators of G protein signaling
Author(s) -
Cappell Steven,
Skowyra Dorota,
Dohlman Henrik
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb167
Subject(s) - skp1 , heterotrimeric g protein , biology , cullin , ubiquitin ligase , f box protein , ubiquitin conjugating enzyme , signal transduction , ubiquitin , gene , protein degradation , microbiology and biotechnology , genetics , g protein
The yeast pheromone pathway consists of a canonical heterotrimeric G protein and MAP kinase cascade. To identify new signaling components we systematically evaluated 870 essential genes using a library of repressible‐promoter strains. Quantitative transcription‐reporter and MAPK activity assays were used to identify strains that exhibit altered pheromone sensitivity. Of the 92 newly identified essential genes required for proper G protein signaling, those involved with protein degradation were most highly‐represented. Included in this group are members of the SCF (Skp‐Cullin‐F‐Box) ubiquitin ligase complex. Further genetic and biochemical analysis reveals that SCF Cdc4 acts together with the Cdc34 ubiquitin conjugating enzyme at the level of the G protein, promotes degradation of the G protein α subunit, Gpa1, in vivo and catalyzes Gpa1 ubiquitination in vitro . These new insights to the G protein signaling network reveal the essential‐genome as an untapped resource for identifying new components and regulators of signal transduction pathways.