Regulation of Homologous Recombination in Human Cells

Project: Research project

Description

DESCRIPTION (provided by applicant): Homologous recombination (HR) is an essential mechanism that maintains genomic integrity in cells by repairing double-strand breaks and repairing damaged replication forks. The RAD51 recombinase catalyzes the central step in the HR pathway, forming joint molecules through homology-dependent strand invasion. The BLM helicase, which is the gene mutated in Bloom's syndrome (BS), regulates RAD51. BLM can promote RAD51 function, for example by generating substrate for RAD51 binding, or it can inhibit RAD51 function by unwinding the products of RAD51 catalysis, which prevents the accumulation of toxic recombination intermediates. Thus, through its regulation of RAD51, BLM has both pro- and anti-recombinogenic functions in HR. We have shown that BLM is modified by small ubiquitin-related modifiers (SUMOs) and that BLM SUMOylation regulates BLM's functions at stalled replication forks, stimulating HR repair. In addition, RAD51 is SUMO-binding, and BLM SUMOylation stimulates BLM's interaction with RAD51 in vitro. Our findings support a model in which BLM SUMOylation controls a switch between BLM's anti- recombinogenic and pro-recombinogenic functions. To test this hypothesis, we have four specific aims: (1) characterization of the role of BLM SUMOylation in stabilization of replication forks;(2) characterization of the role of RAD51 SUMO binding in replication fork stability and HR repair;(3) analysis of the effects of BLM SUMOylation on BLM's biochemical activities and RAD51 function;and (4) analysis of the effects of BLM SUMOylation and RAD51 SUMO binding in genomic integrity. Our studies will elucidate the molecular mechanisms that regulate HR at damaged replication forks, providing insights into the dynamic functions of BLM and RAD51 in HR repair and leading to a deeper understanding of how these functions are regulated by the SUMO pathway. Because HR repair mechanisms are often dysregulated in cancer cells, our work will lead to a better understanding of genomic instability in cancer and will facilitate the exploitation of this instability in cancer treatments. PUBLIC HEALTH RELEVANCE: The homologous recombination pathway repairs DNA damage that arises during the synthesis of DNA. Unregulated homologous recombination causes problems during repair of DNA synthesis failures, and it generates chromosome instability. Chromosome instability is a common feature of cancer cells, and it speeds the development of cancer. The purpose of our research is to characterize the molecular mechanisms that regulate homologous recombination in human cells, which will help in the identification of therapeutic targets for cancer treatments.
StatusFinished
Effective start/end date6/1/113/31/16

Funding

  • National Institutes of Health: $306,229.00
  • National Institutes of Health: $312,808.00
  • National Institutes of Health: $325,775.00
  • National Institutes of Health: $302,216.00
  • National Institutes of Health: $325,775.00

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Sumoylation
Homologous Recombination
Recombinational DNA Repair
Ubiquitin
Neoplasms
Bloom Syndrome
Recombinases
Genomic Instability
Poisons
Catalysis
Genetic Recombination
Joints
Chromosomal Instability
Genes

ASJC

  • Medicine(all)