Systematic proteomics of endogenous human cohesin reveals an interaction with diverse splicing factors and RNA-binding proteins required for mitotic progression.

Abstract

The cohesin complex regulates sister chromatid cohesion, chromosome organization, gene expression, and DNA repair. Cohesin is a ring complex composed of four core subunits and seven regulatory subunits. In an effort to comprehensively identify additional cohesin-interacting proteins, we used gene editing to introduce a dual epitope tag into the endogenous allele of each of 11 known components of cohesin in cultured human cells, and we performed MS analyses on dual-affinity purifications. In addition to reciprocally identifying all known components of cohesin, we found that cohesin interacts with a panoply of splicing factors and RNA-binding proteins (RBPs). These included diverse components of the U4/U6.U5 tri-small nuclear ribonucleoprotein complex and several splicing factors that are commonly mutated in cancer. The interaction between cohesin and splicing factors/RBPs was RNA- and DNA-independent, occurred in chromatin, was enhanced during mitosis, and required RAD21. Furthermore, cohesin-interacting splicing factors and RBPs followed the cohesin cycle and prophase pathway of cell cycle-regulated interactions with chromatin. Depletion of cohesin-interacting splicing factors and RBPs resulted in aberrant mitotic progression. These results provide a comprehensive view of the endogenous human cohesin interactome and identify splicing factors and RBPs as functionally significant cohesin-interacting proteins.

• Authors: Jung-Sik Kim, Xiaoyuan He, Jie Liu, Zhijun Duan, Taeyeon Kim, Julia Gerard, Brian Kim, Manoj M Pillai, William S Lane, William S Noble, Bogdan Budnik, Todd Waldman
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 294 Issue 22 Pg. 8760-8772 (05 31 2019) ISSN: 1083-351X [Electronic] United States
• PMID: 31010829 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
• Copyright: © 2019 Kim et al.

Chemical References

• Cell Cycle Proteins
• Chromatin
• Chromosomal Proteins, Non-Histone
• DNA-Binding Proteins
• RAD21 protein, human
• RNA Splicing Factors
• RNA, Small Interfering
• RNA-Binding Proteins

Topics

• Cell Cycle Proteins (antagonists & inhibitors, genetics, metabolism)
• Cell Line, Tumor
• Chromatin (metabolism)
• Chromosomal Proteins, Non-Histone (genetics, metabolism)
• DNA-Binding Proteins (antagonists & inhibitors, genetics, metabolism)
• Humans
• Microscopy, Fluorescence
• Mitosis
• Protein Binding
• Protein Interaction Maps
• Proteomics
• RNA Interference
• RNA Splicing Factors (antagonists & inhibitors, genetics, metabolism)
• RNA, Small Interfering (metabolism)
• RNA-Binding Proteins (antagonists & inhibitors, genetics, metabolism)
• Cohesins