Understanding the antagonism of retinoblastoma protein dephosphorylation by PNUTS provides insights into the PP1 regulatory code

Meng S Choy, Martina Hieke, Ganesan Senthil Kumar, Greyson R Lewis, Kristofer R Gonzalez-DeWhitt, Rene P Kessler, Benjamin J Stein, Manuel Hessenberger, Angus C Nairn, Wolfgang Peti, Rebecca Page

Research output: Journal article (peer-reviewed)Journal article

78 Citations (Scopus)

Abstract

The serine/threonine protein phosphatase 1 (PP1) dephosphorylates hundreds of key biological targets by associating with nearly 200 regulatory proteins to form highly specific holoenzymes. However, how these proteins direct PP1 specificity and the ability to predict how these PP1 interacting proteins bind PP1 from sequence alone is still missing. PP1 nuclear targeting subunit (PNUTS) is a PP1 targeting protein that, with PP1, plays a central role in the nucleus, where it regulates chromatin decondensation, RNA processing, and the phosphorylation state of fundamental cell cycle proteins, including the retinoblastoma protein (Rb), p53, and MDM2. The molecular function of PNUTS in these processes is completely unknown. Here, we show that PNUTS, which is intrinsically disordered in its free form, interacts strongly with PP1 in a highly extended manner. Unexpectedly, PNUTS blocks one of PP1's substrate binding grooves while leaving the active site accessible. This interaction site, which we have named the arginine site, allowed us to define unique PP1 binding motifs, which advances our ability to predict how more than a quarter of the known PP1 regulators bind PP1. Additionally, the structure shows how PNUTS inhibits the PP1-mediated dephosphorylation of critical substrates, especially Rb, by blocking their binding sites on PP1, insights that are providing strategies for selectively enhancing Rb activity.

Original languageEnglish
Pages (from-to)4097-4102
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume111
Issue number11
DOIs
Publication statusPublished - 18 Mar 2014
Externally publishedYes

Keywords

  • Amino Acid Sequence
  • Calorimetry
  • Chromatin Assembly and Disassembly/physiology
  • Cloning, Molecular
  • Computational Biology
  • Crystallization
  • DNA-Binding Proteins/chemistry
  • Gene Expression Regulation, Enzymologic/genetics
  • Humans
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Sequence Data
  • Nuclear Proteins/chemistry
  • Phosphorylation
  • Protein Conformation
  • Protein Interaction Domains and Motifs/genetics
  • Protein Phosphatase 1/chemistry
  • Proto-Oncogene Proteins c-mdm2/metabolism
  • RNA-Binding Proteins/chemistry
  • Retinoblastoma Protein/metabolism
  • Sequence Alignment
  • Substrate Specificity

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