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

Abdenour Soufi, Peter Noy, M Buckle, A Sawasdichai, Kevin Gaston, Padma-Sheela Jayaraman

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)
169 Downloads (Pure)

Abstract

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 β 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.
Original languageEnglish
Pages (from-to)3288-300
Number of pages13
JournalNucleic Acids Research
Volume37
Issue number10
DOIs
Publication statusPublished - 1 Jun 2009

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