Casein kinase II-mediated phosphorylation of the C terminus of Sp1 decreases its DNA binding activity.
We have previously observed that Sp1, a ubiquitous zinc finger transcription factor, is phosphorylated during terminal differentiation in the whole animal, and this results in decreased DNA binding activity (Leggett, R. W., Armstrong, S. A., Barry, D., and Mueller, C. R. (1995) J. Biol. Chem. 270, 25879-25884). In this study, we demonstrate that casein kinase II (CKII) is able to phosphorylate the C terminus of Sp1 and results in a decrease in DNA binding activity. This suggests that CKII may be responsible for the observed regulation of Sp1. Mutation of a consensus CKII site at amino acid 579, within the second zinc finger, eliminates phosphorylation of this site and the CKII-mediated inhibition of Sp1 binding. Phosphopeptide analysis confirms the presence of a CKII site at Thr-579 as well as additional sites within the C terminus. No gross changes in CKII subunit levels were seen during de-differentiation associated with liver regeneration. The serine/threonine phosphatase PP1 was identified as the endogenous liver nuclear protein able to dephosphorylate Sp1 but again no gross changes in activity were observed in the regenerating liver. Okadaic acid treatment of K562 cells increases Sp1 phosphorylation and inhibits its DNA binding activity suggesting that steady state levels of Sp1 phosphorylation are established by a balance between kinase and phosphatase activities.
Armstrong
S A
SA
Cancer Research Laboratories, Queen's University, Kingston, Ontario K7L 3N6, Canada.
Barry
D A
DA
Leggett
R W
RW
Mueller
C R
CR
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D047390:Casein Kinase II; D002454:Cell Differentiation; D004247:DNA / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008115:Liver Regeneration; D008297:Male; D008969:Molecular Sequence Data; D019319:Okadaic Acid / Q000494:pharmacology; D010766:Phosphorylation; D017354:Point Mutation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051381:Rats; D016329:Sp1 Transcription Factor / Q000235:genetics / Q000378:metabolism*
D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D016329:Sp1 Transcription Factor; D019319:Okadaic Acid; D004247:DNA; D047390:Casein Kinase II; D017346:Protein Serine-Threonine Kinases