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Title: The Histone Modification Domain of Paf1 Complex Subunit Rtf1 Directly Stimulates H2B Ubiquitylation through an Interaction with Rad6

The five-subunit yeast Paf1 Complex (Paf1C) regulates all stages of transcription and is critical for the monoubiquitylation of histone H2B (H2Bub), a modification that broadly influences chromatin structure and eukaryotic transcription. Here we show that the histone modification domain (HMD) of Paf1C subunit Rtf1 directly interacts with the ubiquitin conjugase Rad6 and stimulates H2Bub independently of transcription. We present the crystal structure of the Rtf1 HMD and use site-specific, in vivo crosslinking to identify a conserved Rad6 interaction surface. Utilizing ChIP-exo analysis, we define the localization patterns of the H2Bub machinery at high resolution and demonstrate the importance of Paf1C in targeting the Rtf1 HMD, and thereby H2Bub, to its appropriate genomic locations. Finally, we observe HMD-dependent stimulation of H2Bub in a transcription-free, reconstituted in vitro system. Taken together, our results argue for an active role for Paf1C in promoting H2Bub and ensuring its proper localization in vivo.
Authors:
 [1] ;  [1] ;  [2] ;  [1] ;  [3] ;  [2] ;  [4] ;  [1] ;  [5] ;  [6] ;  [6] ;  [1] ;  [2] ;  [1]
  1. Univ. of Pittsburgh, PA (United States)
  2. Pennsylvania State Univ., University Park, PA (United States)
  3. Pennsylvania State Univ., University Park, PA (United States); Southern Medical Univ., Guangzhou (China)
  4. Univ. of Pittsburgh School of Medicine, PA (United States)
  5. Brookhaven National Lab. (BNL), Upton, NY (United States)
  6. Korea Advanced Inst. Science and Technology (KAIST), Daejeon (Korea, Republic of)
Publication Date:
Report Number(s):
BNL-113612-2017-JA
Journal ID: ISSN 1097-2765
Grant/Contract Number:
SC00112704
Type:
Accepted Manuscript
Journal Name:
Molecular Cell
Additional Journal Information:
Journal Volume: 64; Journal Issue: 4; Journal ID: ISSN 1097-2765
Publisher:
Elsevier - Cell Press
Research Org:
Brookhaven National Laboratory (BNL), Upton, NY (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES
OSTI Identifier:
1347296