Splicing function of mitotic regulators links R-loop-mediated DNA damage to tumor cell killing

Yihan Wan; Xiaobin Zheng; Haiyang Chen; Yuxuan Guo; Hao Jiang; Xiaonan He; Xueliang Zhu; Yixian Zheng

doi:10.1083/jcb.201409073

Splicing function of mitotic regulators links R-loop-mediated DNA damage to tumor cell killing

J Cell Biol. 2015 Apr 27;209(2):235-46. doi: 10.1083/jcb.201409073.

Authors

Yihan Wan¹, Xiaobin Zheng², Haiyang Chen², Yuxuan Guo², Hao Jiang¹, Xiaonan He³, Xueliang Zhu⁴, Yixian Zheng⁵

Affiliations

¹ State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China Department of Embryology, Carnegie Institution for Science, Baltimore, MD 21218.
² Department of Embryology, Carnegie Institution for Science, Baltimore, MD 21218.
³ State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
⁴ State Key Laboratory of Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China zheng@ciwemb.edu xlzhu@sibcb.ac.cn.
⁵ Department of Embryology, Carnegie Institution for Science, Baltimore, MD 21218 zheng@ciwemb.edu xlzhu@sibcb.ac.cn.

Abstract

Although studies suggest that perturbing mitotic progression leads to DNA damage and p53 activation, which in turn lead to either cell apoptosis or senescence, it remains unclear how mitotic defects trigger p53 activation. We show that BuGZ and Bub3, which are two mitotic regulators localized in the interphase nucleus, interact with the splicing machinery and are required for pre-mRNA splicing. Similar to inhibition of RNA splicing by pladienolide B, depletion of either BuGZ or Bub3 led to increased formation of RNA-DNA hybrids (R-loops), which led to DNA damage and p53 activation in both human tumor cells and primary cells. Thus, R-loop-mediated DNA damage and p53 activation offer a mechanistic explanation for apoptosis of cancer cells and senescence of primary cells upon disruption of the dual-function mitotic regulators. This demonstrates the importance of understanding the full range of functions of mitotic regulators to develop antitumor drugs.

Publication types

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

MeSH terms

Animals
Apoptosis*
Biomarkers / analysis
Blotting, Western
Cell Cycle Proteins / genetics
Cell Cycle Proteins / metabolism*
Cellular Senescence
DNA Damage*
Drosophila melanogaster / metabolism
Fibroblasts / cytology
Fibroblasts / metabolism
Fluorescent Antibody Technique
High-Throughput Nucleotide Sequencing
Humans
Immunoprecipitation
Microtubule-Associated Proteins / genetics
Microtubule-Associated Proteins / metabolism*
Mitosis / physiology
Neoplasms / genetics
Neoplasms / metabolism
Neoplasms / pathology*
Poly-ADP-Ribose Binding Proteins
RNA Splicing*
RNA, Messenger / genetics
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Tumor Cells, Cultured
Tumor Suppressor Protein p53 / genetics
Tumor Suppressor Protein p53 / metabolism*

Substances

BUB3 protein, human
Biomarkers
Cell Cycle Proteins
Microtubule-Associated Proteins
Poly-ADP-Ribose Binding Proteins
RNA, Messenger
TP53 protein, human
Tumor Suppressor Protein p53
ZNF207 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding