The ubiquitin hydrolase USP22 contributes to 3'-end processing of JAK-STAT-inducible genes

FASEB J. 2012 Feb;26(2):842-54. doi: 10.1096/fj.11-189498. Epub 2011 Nov 8.

Abstract

The JAK-STAT (Janus kinase-signal transducer and activator of transcription) signaling pathway drives cellular growth, differentiation, and the immune response. STAT-activated gene expression is both rapid and transient and requires dynamic post-translational modification of the chromatin template. We previously showed that monoubiquitination of histone H2B (ubH2B) is highly dynamic at the STAT1 target gene, interferon regulatory factor 1 (IRF1), suggesting that a deubiquitinase is recruited during gene activation. Here, we report that RNAi-mediated knockdown of the ubiquitin hydrolase, USP22, results in 2-fold higher ubH2B, and 2-fold lower transcriptional elongation at IRF1. We also demonstrate that USP22 depletion diminishes 3'-end cleavage/polyadenylation by 2- to 3-fold. Furthermore, the polyadenylation factor CPSF73 is not effectively recruited, and serine 2 phosphorylation (Ser2P) of the C-terminal domain of RNA polymerase II is also disrupted. The transcriptional and processing defects observed in the USP22-knockdown cells are reversed by transient USP22 overexpression. Together, these results suggest that ubH2B helps recruit polyadenylation factors to STAT1-activated genes. We propose a working model, wherein a cycle of H2B ubiquitination/deubiquitination specifies Ser2P to regulate elongation and 3'-end processing of JAK-STAT-inducible mRNAs. These results further elaborate USP22 function and its role as a putative cancer stem cell marker.

MeSH terms

  • 3' Untranslated Regions
  • Base Sequence
  • Cell Death / genetics
  • Cell Line
  • Chromatin / genetics
  • Chromatin / metabolism
  • Cleavage And Polyadenylation Specificity Factor / metabolism
  • DNA Primers / genetics
  • Gene Knockdown Techniques
  • Histones / metabolism
  • Humans
  • Interferon Regulatory Factor-1 / genetics
  • Interferon Regulatory Factor-1 / metabolism
  • Janus Kinases / metabolism*
  • Models, Biological
  • Nuclear Proteins / metabolism
  • Protein Processing, Post-Translational
  • RNA 3' End Processing*
  • RNA Polymerase II / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • STAT Transcription Factors / metabolism*
  • Signal Transduction / genetics
  • Signal Transduction / physiology
  • Thiolester Hydrolases / antagonists & inhibitors
  • Thiolester Hydrolases / genetics
  • Thiolester Hydrolases / metabolism*
  • Transcription Factors
  • Ubiquitin Thiolesterase
  • Ubiquitination

Substances

  • 3' Untranslated Regions
  • Chromatin
  • Cleavage And Polyadenylation Specificity Factor
  • DNA Primers
  • Histones
  • IRF1 protein, human
  • Interferon Regulatory Factor-1
  • Nuclear Proteins
  • PAF1 protein, human
  • RNA, Messenger
  • STAT Transcription Factors
  • Transcription Factors
  • Janus Kinases
  • RNA Polymerase II
  • Thiolester Hydrolases
  • Ubiquitin Thiolesterase
  • Usp22 protein, human