title: Chicken progesterone receptor is phosphorylated by a DNA-dependent protein kinase during in vitro transcription assays.
abstract: We have reported previously that chicken progesterone receptor (PR) is phosphorylated in vivo in response to progesterone administration. Three phosphorylation sites have been reported, two of which show increased phosphorylation in response to hormone and one which is phosphorylated only in response to hormone administration. We found previously that PR lacking the hormone-dependent phosphorylation is active in an in vitro transcription assay. Since the source of general transcription factors is a HeLa nuclear extract which contains many kinases, we have analyzed the receptor for phosphorylation during the in vitro transcription assay. We report here that the receptor is rapidly and efficiently phosphorylated on new sites, causing a change in receptor mobility on sodium dodecyl sulfate-gels. This phosphorylation is strictly dependent upon the presence of double stranded DNA. A DNA-activated protein kinase with similar properties has been isolated previously from HeLa cell nuclei. We find that phosphorylation of PR with this purified enzyme mimics the phosphorylation observed in the transcription assay. These data suggest that a previously undetected additional series of DNA-dependent phosphorylations may be required for activation of the PR.
authors.affiliation: Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
authors.forename: N L
authors.initials: NL
authors.lastname: Weigel
authors.forename: T H
authors.initials: TH
authors.lastname: Carter
authors.forename: W T
authors.initials: WT
authors.lastname: Schrader
authors.forename: B W
authors.initials: BW
authors.lastname: O'Malley
mesh_terms: D000818:Animals; D001345:Autoradiography; D002645:Chickens / Q000502:physiology*; D004247:DNA / Q000502:physiology; D015151:Immunoblotting; D066298:In Vitro Techniques; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D011980:Receptors, Progesterone / Q000378:metabolism*; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: D011980:Receptors, Progesterone; D004247:DNA; D011494:Protein Kinases