Transcription-replication conflicts as a source of common fragile site instability caused by BMI1-RNF2 deficiency


Autoři: Anthony Sanchez aff001;  Angelo de Vivo aff001;  Peter Tonzi aff002;  Jeonghyeon Kim aff001;  Tony T. Huang aff002;  Younghoon Kee aff001
Působiště autorů: Department of Cell Biology, Microbiology, and Molecular Biology, College of Arts and Sciences, University of South Florida, Tampa, Florida, United States of America aff001;  Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, New York, New York, United States of America aff002
Vyšlo v časopise: Transcription-replication conflicts as a source of common fragile site instability caused by BMI1-RNF2 deficiency. PLoS Genet 16(3): e32767. doi:10.1371/journal.pgen.1008524
Kategorie: Research Article
doi: 10.1371/journal.pgen.1008524

Souhrn

Common fragile sites (CFSs) are breakage-prone genomic loci, and are considered to be hotspots for genomic rearrangements frequently observed in cancers. Understanding the underlying mechanisms for CFS instability will lead to better insight on cancer etiology. Here we show that Polycomb group proteins BMI1 and RNF2 are suppressors of transcription-replication conflicts (TRCs) and CFS instability. Cells depleted of BMI1 or RNF2 showed slower replication forks and elevated fork stalling. These phenotypes are associated with increase occupancy of RNA Pol II (RNAPII) at CFSs, suggesting that the BMI1-RNF2 complex regulate RNAPII elongation at these fragile regions. Using proximity ligase assays, we showed that depleting BMI1 or RNF2 causes increased associations between RNAPII with EdU-labeled nascent forks and replisomes, suggesting increased TRC incidences. Increased occupancy of a fork protective factor FANCD2 and R-loop resolvase RNH1 at CFSs are observed in RNF2 CRISPR-KO cells, which are consistent with increased transcription-associated replication stress in RNF2-deficient cells. Depleting FANCD2 or FANCI proteins further increased genomic instability and cell death of the RNF2-deficient cells, suggesting that in the absence of RNF2, cells depend on these fork-protective factors for survival. These data suggest that the Polycomb proteins have non-canonical roles in suppressing TRC and preserving genomic integrity.

Klíčová slova:

Anemia – Cyclins – DNA replication – DNA transcription – Genetic networks – Immunoprecipitation – Protein structure networks – Small interfering RNAs


Zdroje

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