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Structural basis for the multi-activity factor Rad5 in replication stress tolerance.

Miaomiao Shen1,2,3, Nalini Dhingra4, Quan Wang5

  • 1Department of Biochemistry and Molecular Biology, Tianjin Medical University, 300070, Tianjin, P. R. China.

Nature Communications
|January 13, 2021
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Summary
This summary is machine-generated.

The yeast protein Rad5, crucial for DNA repair and cell survival, has its structure revealed. Its HIRAN domain plays multiple roles in DNA metabolism and replication stress tolerance.

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Area of Science:

  • Molecular Biology
  • Structural Biology
  • Genetics

Background:

  • Rad5 protein and its orthologs are vital for eukaryotic replication stress tolerance and cell survival.
  • These proteins possess multiple activities, including ubiquitin ligase, replication fork remodeling, and DNA lesion targeting.

Purpose of the Study:

  • To present the crystal structure of a nearly full-length yeast Rad5 protein.
  • To elucidate the functional roles of Rad5's domains, particularly the HIRAN domain, in DNA metabolism and replication stress tolerance.

Main Methods:

  • X-ray crystallography to determine the structure of Rad5.
  • Biochemical assays to investigate protein-DNA and protein-protein interactions.
  • Cellular studies to assess Rad5 function in vivo.

Main Results:

  • The crystal structure reveals three distinct, interconnected domains within Rad5.
  • The HIRAN domain interacts with PCNA (Proliferating Cell Nuclear Antigen), mediates poly-ubiquitination, binds DNA, and contributes to replication fork regression.
  • These findings define a novel multifunctional type of HIRAN domain.

Conclusions:

  • The structure of Rad5 provides insights into how its domains coordinate for autonomous and integrated activities.
  • The multifunctional HIRAN domain is key to Rad5's role in replication stress tolerance.
  • This work offers a framework for understanding Rad5's integrated functions in maintaining genomic stability.