H3K36 Methylation and the Chromodomain Protein Eaf3 Are Required for Proper Cotranscriptional Spliceosome Assembly

Calvin S Leung; Stephen M Douglass; Marco Morselli; Matthew B Obusan; Marat S Pavlyukov; Matteo Pellegrini; Tracy L Johnson

doi:10.1016/j.celrep.2019.05.100

H3K36 Methylation and the Chromodomain Protein Eaf3 Are Required for Proper Cotranscriptional Spliceosome Assembly

Cell Rep. 2019 Jun 25;27(13):3760-3769.e4. doi: 10.1016/j.celrep.2019.05.100.

Authors

Calvin S Leung¹, Stephen M Douglass², Marco Morselli³, Matthew B Obusan², Marat S Pavlyukov⁴, Matteo Pellegrini⁵, Tracy L Johnson⁶

Affiliations

¹ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA.
² Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA.
³ Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Institute for Genomics and Proteomics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Institute for Quantitative and Computational Biosciences, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁴ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russian Federation.
⁵ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA; Institute for Genomics and Proteomics, University of California, Los Angeles, Los Angeles, CA 90095, USA; Institute for Quantitative and Computational Biosciences, University of California, Los Angeles, Los Angeles, CA 90095, USA.
⁶ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA 90095, USA; Department of Molecular, Cell, and Developmental Biology, University of California, Los Angeles, Los Angeles, CA 90095, USA. Electronic address: tljohnson@ucla.edu.

Abstract

In the eukaryotic cell, spliceosomes assemble onto pre-mRNA cotranscriptionally. Spliceosome assembly takes place in the context of the chromatin environment, suggesting that the state of the chromatin may affect splicing. The molecular details and mechanisms through which chromatin affects splicing, however, are still unclear. Here, we show a role for the histone methyltransferase Set2 and its histone modification, H3K36 methylation, in pre-mRNA splicing through high-throughput sequencing. Moreover, the effect of H3K36 methylation on pre-mRNA splicing is mediated through the chromodomain protein Eaf3. We find that Eaf3 is recruited to intron-containing genes and that Eaf3 interacts with the splicing factor Prp45. Eaf3 acts with Prp45 and Prp19 after formation of the precatalytic B complex around the time of splicing activation, thus revealing the step in splicing that is regulated by H3K36 methylation. These studies support a model whereby H3K36 facilitates recruitment of an "adapter protein" to support efficient, constitutive splicing.

Keywords: Eaf3; H3K36; Set2; chromatin; cotranscriptional splicing; methylation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Acetyltransferases / genetics
Acetyltransferases / metabolism*
Histones / genetics
Histones / metabolism*
Methylation
RNA Splicing Factors / genetics
RNA Splicing Factors / metabolism
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / metabolism*
Saccharomyces cerevisiae Proteins / genetics
Saccharomyces cerevisiae Proteins / metabolism*
Spliceosomes / genetics
Spliceosomes / metabolism*
Transcription, Genetic*

Substances

Histones
PRP19 protein, S cerevisiae
PRP45 protein, S cerevisiae
RNA Splicing Factors
Saccharomyces cerevisiae Proteins
Acetyltransferases
Eaf3 protein, S cerevisiae

Abstract

Publication types

MeSH terms

Substances

Grants and funding