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CK2 phosphorylation of the PRH/Hex homeodomain functions as a reversible switch for DNA binding.

TitleCK2 phosphorylation of the PRH/Hex homeodomain functions as a reversible switch for DNA binding.
Publication TypeJournal Article
Year of Publication2009
AuthorsSoufi A, Noy P, Buckle M, Sawasdichai A, Gaston K, Jayaraman P-S
JournalNucleic Acids Res
Date Published2009 Jun
KeywordsAmino Acid Sequence, Casein Kinase II, DNA, Homeodomain Proteins, Humans, K562 Cells, Molecular Sequence Data, Mutation, Phosphoproteins, Phosphorylation, Protein Structure, Tertiary, Repressor Proteins, Serine, Transcription Factors

The proline-rich homeodomain protein (PRH/Hex) regulates transcription by binding to specific DNA sequences and regulates mRNA transport by binding to translation initiation factor eIF4E. Protein kinase CK2 plays multiple roles in the regulation of gene expression and cell proliferation. Here, we show that PRH interacts with the beta subunit of CK2 in vitro and in cells and that CK2 phosphorylates PRH. Phosphorylation of PRH by CK2 inhibits the DNA binding activity of this protein and dephosphorylation restores DNA binding indicating that this modification acts as a reversible switch. We show that phosphorylation of the homeodomain is sufficient to block DNA binding and we identify two amino acids within this the domain that are phosphorylated by CK2: S163 and S177. Site-directed mutagenesis demonstrates that mutation of either of these residues to glutamic acid partially mimics phosphorylation but is insufficient to completely block DNA binding whereas an S163E/S177E double mutation severely inhibits DNA binding. Significantly, the S163E and S177E mutations and the S163E/S177E double mutation all inhibit the ability of PRH to regulate transcription in cells. Since these amino acids are conserved between many homeodomain proteins, our results suggest that CK2 may regulate the activity of several homeodomain proteins in this manner.

Alternate JournalNucleic Acids Res.
PubMed ID19324893
PubMed Central IDPMC2691835
Grant List / / Biotechnology and Biological Sciences Research Council / United Kingdom
/ / Wellcome Trust / United Kingdom
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