Hormonal regulation and identification of chicken progesterone receptor phosphorylation sites

Larry Denner, William T. Schrader, Bert W. O'Malley, Nancy L. Weigel

Research output: Contribution to journalArticle

97 Citations (Scopus)

Abstract

The present studies examine the effects of in vivo and in situ progesterone treatment in the regulation of site-specific phosphorylation of the chicken oviduct progesterone receptor (PR). By gas-phase protein sequencing we have identified three hormonally regulated phosphorylation sites: Ser-211, Ser-260, and Ser-530. We determined phosphorylation stoichiometries by analyzing the amounts of phosphorylated and dephosphorylated serine at each site. Stoichiometries of sites 211 and 260 were about 20% under basal conditions and increased 1.5-2-fold by in situ progesterone treatment. Site 530 was virtually absent under basal conditions and induced to >33% by in situ progesterone treatment. We tested several protein kinases for phosphorylation of the PR in vitro on these sites or peptides containing these sites. We found that the catalytic subunit of cAMP-dependent protein kinase mimicked the in vivo, hormone-induced altered mobility of the PRs in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Both the in vivo and in vitro alterations were reversed by alkaline phosphatase. Finally, we showed that cAMP-dependent protein kinase phosphorylated Ser-528.

Original languageEnglish (US)
Pages (from-to)16548-16555
Number of pages8
JournalJournal of Biological Chemistry
Volume265
Issue number27
StatePublished - Sep 25 1990
Externally publishedYes

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Phosphorylation
Progesterone Receptors
Chickens
Progesterone
Cyclic AMP-Dependent Protein Kinases
Stoichiometry
Oviducts
Protein Sequence Analysis
Electrophoresis
Sodium Dodecyl Sulfate
Protein Kinases
Serine
Alkaline Phosphatase
Polyacrylamide Gel Electrophoresis
Catalytic Domain
Gases
Hormones
Peptides
Proteins
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Hormonal regulation and identification of chicken progesterone receptor phosphorylation sites. / Denner, Larry; Schrader, William T.; O'Malley, Bert W.; Weigel, Nancy L.

In: Journal of Biological Chemistry, Vol. 265, No. 27, 25.09.1990, p. 16548-16555.

Research output: Contribution to journalArticle

Denner, Larry ; Schrader, William T. ; O'Malley, Bert W. ; Weigel, Nancy L. / Hormonal regulation and identification of chicken progesterone receptor phosphorylation sites. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 27. pp. 16548-16555.
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