Interaction between transmembrane domains five and six of the α-factor receptor

Peter H Dube, Anthony DeCostanzo, James B. Konopka

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

The α-factor pheromone receptor (STE2) activates a G protein signal pathway that induces conjugation of the yeast Saccharomyces cerevisiae. Previous studies implicared the third intracellular loop of this receptor in G protein activation. Therefore, the roles of transmembrane domains five and six (TMD5 and -6) that bracket the third intracellular loop were analyzed by scanning mutagenesis in which each residue was substituted with cysteine. Out of 42 mutants examined, four constitutive mutants and two strong loss-of-function mutants were identified. Double mutants combining Cys substitutions in TMD5 and TMD6 gave a broader range of phenotypes. Interestingly, a V223C mutation in TMD5 caused constitutive activity when combined with the L247C, L248C, or S251C mutations in TMD6. Also, the L226C mutation in TMD5 caused constitutive activity when combined with either the M250C or S251C mutations in TMD6. The residues affected by these mutations are predicted to fall on one side of their respective helices, suggesting that they may interact. In support of this, cysteines substituted at position 223 in TMD5 and position 247 in TMD6 formed a disulfide bond, providing the first direct evidence of an interaction between these transmembrane domains in the α-factor receptor. Altogether, these results identify an important region of interaction between conserved hydrophobic regions at the base of TMD5 and TMD6 that is required for the proper regulation of receptor signaling.

Original languageEnglish (US)
Pages (from-to)26492-26499
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number34
DOIs
StatePublished - Aug 25 2000
Externally publishedYes

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GTP-Binding Proteins
Yeast
Cysteine
Pheromone Receptors
Mutagenesis
Mutation
Disulfides
Substitution reactions
Chemical activation
Scanning
Saccharomyces cerevisiae
Signal Transduction
Yeasts
Phenotype

ASJC Scopus subject areas

  • Biochemistry

Cite this

Interaction between transmembrane domains five and six of the α-factor receptor. / Dube, Peter H; DeCostanzo, Anthony; Konopka, James B.

In: Journal of Biological Chemistry, Vol. 275, No. 34, 25.08.2000, p. 26492-26499.

Research output: Contribution to journalArticle

Dube, Peter H ; DeCostanzo, Anthony ; Konopka, James B. / Interaction between transmembrane domains five and six of the α-factor receptor. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 34. pp. 26492-26499.
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