Phosphorylation regulates the Star-PAP-PIPKIα interaction and directs specificity toward mRNA targets

Nucleic Acids Res. 2015 Aug 18;43(14):7005-20. doi: 10.1093/nar/gkv676. Epub 2015 Jul 2.

Abstract

Star-PAP is a nuclear non-canonical poly(A) polymerase (PAP) that shows specificity toward mRNA targets. Star-PAP activity is stimulated by lipid messenger phosphatidyl inositol 4,5 bisphoshate (PI4,5P2) and is regulated by the associated Type I phosphatidylinositol-4-phosphate 5-kinase that synthesizes PI4,5P2 as well as protein kinases. These associated kinases act as coactivators of Star-PAP that regulates its activity and specificity toward mRNAs, yet the mechanism of control of these interactions are not defined. We identified a phosphorylated residue (serine 6, S6) on Star-PAP in the zinc finger region, the domain required for PIPKIα interaction. We show that S6 is phosphorylated by CKIα within the nucleus which is required for Star-PAP nuclear retention and interaction with PIPKIα. Unlike the CKIα mediated phosphorylation at the catalytic domain, Star-PAP S6 phosphorylation is insensitive to oxidative stress suggesting a signal mediated regulation of CKIα activity. S6 phosphorylation together with coactivator PIPKIα controlled select subset of Star-PAP target messages by regulating Star-PAP-mRNA association. Our results establish a novel role for phosphorylation in determining Star-PAP target mRNA specificity and regulation of 3'-end processing.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Casein Kinase I / metabolism
  • Cell Nucleus / enzymology
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Nucleotidyltransferases
  • Oxidative Stress
  • Phosphatidylinositol 4,5-Diphosphate / metabolism
  • Phosphorylation
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism*
  • Polynucleotide Adenylyltransferase / chemistry
  • Polynucleotide Adenylyltransferase / metabolism*
  • Protein Binding
  • RNA 3' End Processing*
  • RNA, Messenger / metabolism*
  • Serine / metabolism
  • Signal Transduction
  • Zinc Fingers

Substances

  • Phosphatidylinositol 4,5-Diphosphate
  • RNA, Messenger
  • Serine
  • Phosphotransferases (Alcohol Group Acceptor)
  • 1-phosphatidylinositol-4-phosphate 5-kinase
  • Casein Kinase I
  • Nucleotidyltransferases
  • TUT1 protein, human
  • Polynucleotide Adenylyltransferase