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The Histone Modification Domain of Paf1 Complex Subunit Rtf1 Directly Stimulates H2B Ubiquitylation through an Interaction with Rad6.
- Source :
-
Molecular cell [Mol Cell] 2016 Nov 17; Vol. 64 (4), pp. 815-825. Date of Electronic Publication: 2016 Nov 10. - Publication Year :
- 2016
-
Abstract
- 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.<br /> (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Subjects :
- Amino Acid Motifs
Binding Sites
Chromosomal Proteins, Non-Histone genetics
Chromosomal Proteins, Non-Histone metabolism
Cross-Linking Reagents chemistry
Crystallography, X-Ray
Formaldehyde chemistry
Histones chemistry
Histones genetics
Models, Molecular
Nuclear Proteins genetics
Nuclear Proteins metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Interaction Domains and Motifs
RNA Polymerase II genetics
RNA Polymerase II metabolism
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins chemistry
Saccharomyces cerevisiae Proteins genetics
TATA-Box Binding Protein chemistry
TATA-Box Binding Protein genetics
Transcription, Genetic
Transcriptional Elongation Factors genetics
Transcriptional Elongation Factors metabolism
Ubiquitin-Conjugating Enzymes chemistry
Ubiquitin-Conjugating Enzymes genetics
Ubiquitination
Gene Expression Regulation, Fungal
Histones metabolism
Saccharomyces cerevisiae metabolism
Saccharomyces cerevisiae Proteins metabolism
TATA-Box Binding Protein metabolism
Ubiquitin-Conjugating Enzymes metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1097-4164
- Volume :
- 64
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Molecular cell
- Publication Type :
- Academic Journal
- Accession number :
- 27840029
- Full Text :
- https://doi.org/10.1016/j.molcel.2016.10.008