5EMX

Crystal structure of the S. cerevisiae Rtf1 histone modification domain mutant R124A R126A R128A

Summary for 5EMX

• Entry DOI: 10.2210/pdb5emx/pdb
• Related: 5E8B
• Descriptor: RNA polymerase-associated protein RTF1 (2 entities in total)
• Functional Keywords: transcription, paf1, chromatin, rtf1
• Biological source: Saccharomyces cerevisiae (Baker's yeast)
• Cellular location: Nucleus, nucleoplasm : P53064
• Total number of polymer chains: 2
• Total formula weight: 17032.86

Authors

Wier, A.D.,Heroux, A.,VanDemark, A.P. (deposition date: 2015-11-06, release date: 2016-10-26, Last modification date: 2024-03-06)

Primary citation

Van Oss, S.B.,Shirra, M.K.,Bataille, A.R.,Wier, A.D.,Yen, K.,Vinayachandran, V.,Byeon, I.L.,Cucinotta, C.E.,Heroux, A.,Jeon, J.,Kim, J.,VanDemark, A.P.,Pugh, B.F.,Arndt, K.M.
The Histone Modification Domain of Paf1 Complex Subunit Rtf1 Directly Stimulates H2B Ubiquitylation through an Interaction with Rad6.
Mol.Cell, 64:815-825, 2016

PubMed 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.

PubMed: 27840029
DOI: 10.1016/j.molcel.2016.10.008

Experimental method

X-RAY DIFFRACTION (1.399 Å)