Histone and high mobility group protein phosphorylation in the thyroid: regulation by cyclic nucleotides.
A variety of cyclic nucleotide analogs and other agents that affect thyroid cyclic nucleotide metabolism were used to investigate the role of cAMP and cGMP in regulating nuclear protein phosphorylation in calf thyroid slices labeled in vitro with [32P]orthophosphate. Two major groups of acid-soluble proteins were studied. Group I consisted of proteins whose phosphorylation is stimulated by TSH [histones H1 and H3, high mobility group (HMG) protein 14, and the HMG 14/17-like protein PS.3]; group II included representatives of a spectrum of proteins whose phosphorylation is unaffected by TSH (histones H2A, H2B, and H4, HMG 17, the HMG 14/17-like protein PS.2, and the nonhistone protein AS.1). The effects of TSH (50 mU/ml) on the 32P labeling of group I proteins were partially reproduced by (Bu)2cAMP (1 mM), 8-bromo-cAMP (1 mM), and butyrate (2 mM), and closely mimicked by 8-(4-chlorophenylthio)cAMP (1 mM), forskolin (25 microM), and butyrate (10 mM). (Bu)2cGMP (1 mM), 8-bromo-cGMP (1 mM), and carbachol (50 microM) had no effect on protein phosphorylation. NaNO2 (20 mM), which markedly increases cGMP concentration in calf thyroid slices, decreased the 32P labeling of group I proteins and also affected, to varying extents, the phosphorylation of the group II proteins. The phosphodiesterase inhibitor methylisobutylxanthine (0.5 mM) had generally minor effects on 32P labeling; however, it did counteract the effects of NaNO2 on group I protein phosphorylation. Our results provide strong support for the hypothesis that TSH-dependent phosphorylation of group I proteins is mediated by cAMP, but they provide little evidence of cGMP regulation of histone or HMG protein phosphorylation.
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D002217:Carbachol / Q000494:pharmacology; D002417:Cattle; D005576:Colforsin; D004224:Diterpenes / Q000494:pharmacology; D006609:High Mobility Group Proteins / Q000378:metabolism*; D006657:Histones / Q000494:pharmacology*; D009712:Nucleotides, Cyclic / Q000494:pharmacology / Q000502:physiology*; D010766:Phosphorylation; D012977:Sodium Nitrite / Q000494:pharmacology; D013961:Thyroid Gland / Q000378:metabolism*
D004224:Diterpenes; D006609:High Mobility Group Proteins; D006657:Histones; D009712:Nucleotides, Cyclic; D005576:Colforsin; D002217:Carbachol; D012977:Sodium Nitrite; D015056:1-Methyl-3-isobutylxanthine