The protein kinase CK2 phosphorylates SNAP190 to negatively regulate SNAPC DNA binding and human U6 transcription by RNA polymerase III.
Human U6 small nuclear RNA gene transcription by RNA polymerase III requires the general transcription factor SNAP(C), which binds to human small nuclear RNA core promoter elements and nucleates pre-initiation complex assembly with the Brf2-TFIIIB complex. Multiple components in this pathway are phosphorylated by the protein kinase CK2, including the Bdp1 subunit of the Brf2-TFIIIB complex, and RNA polymerase III, with negative and positive outcomes for U6 transcription, respectively. However, a role for CK2 phosphorylation of SNAP(C) in U6 transcription has not been defined. In this report, we investigated the role of CK2 in modulating the transcriptional properties of SNAP(C) and demonstrate that within SNAP(C), CK2 phosphorylates the N-terminal half of the SNAP190 subunit at two regions (amino acids 20-63 and 514-545) that each contain multiple CK2 consensus sites. SNAP190 phosphorylation by CK2 inhibits both SNAP(C) DNA binding and U6 transcription activity. Mutational analyses of SNAP190 support a model wherein CK2 phosphorylation triggers an allosteric inhibition of the SNAP190 Myb DNA binding domain.
Department of Biochemistry & Molecular Biology, Michigan State University, East Lansing, Michigan 48824, USA.
Liping
L
Gu
Rhonda
R
Husain-Ponnampalam
Susanne
S
Hoffmann-Benning
R William
RW
Henry
D000495:Allosteric Site; D047390:Casein Kinase II / Q000378:metabolism*; D002467:Cell Nucleus / Q000378:metabolism; D004247:DNA / Q000737:chemistry; D018076:DNA, Complementary / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D017434:Protein Structure, Tertiary; D012320:RNA Polymerase III / Q000737:chemistry / Q000502:physiology*; D012342:RNA, Small Nuclear / Q000737:chemistry*; D014157:Transcription Factors / Q000737:chemistry*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; D010455:Peptides; D012342:RNA, Small Nuclear; C116351:SNAPC4 protein, human; D014157:Transcription Factors; C405486:U6 small nuclear RNA; D004247:DNA; D047390:Casein Kinase II; D012320:RNA Polymerase III