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Updated: Aug 13, 2026

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells
08:38

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells

Published on: March 3, 2015

The human checkpoint protein hRad17 interacts with the PCNA-like proteins hRad1, hHus1, and hRad9

M Rauen1, M A Burtelow, V M Dufault

  • 1Division of Developmental Oncology Research, Mayo Clinic, Rochester, Minnesota 55902, USA.

The Journal of Biological Chemistry
|July 8, 2000
PubMed

Insights

Human Rad17 (hRad17) interacts with PCNA-like checkpoint proteins hRad1, hRad9, and hHus1. This interaction, crucial for DNA damage response, mirrors the RFC-PCNA clamp-loader relationship.

Area of Science:

  • Cellular biology
  • Molecular genetics
  • DNA repair mechanisms

Background:

  • DNA damage triggers cell cycle checkpoints to halt progression.
  • Yeast DNA damage response genes (rad1, rad9, hus1, rad17) have mammalian homologs.
  • Human Rad1, Rad9, and Hus1 proteins are structurally similar to PCNA (sliding clamp).
  • Rad17 homologs resemble Replication Factor C (RFC) subunits, a PCNA clamp loader.

Purpose of the Study:

  • To investigate the interaction between human Rad17 (hRad17) and PCNA-like checkpoint proteins (hRad1, hRad9, hHus1).
  • To determine if this interaction resembles the established RFC-PCNA clamp-loader relationship.
  • To assess the effect of DNA damage on the hRad17-checkpoint protein association.

Main Methods:

  • Demonstration of endogenous hRad17 interaction with hRad1, hRad9, and hHus1.
  • Mutational analysis of hRad1 and hRad17 to characterize interaction properties.
  • Investigation of DNA damage effects on the association between hRad17 and clamp-like checkpoint proteins.

Main Results:

  • Endogenous hRad17 was shown to interact with hRad1, hRad9, and hHus1.
  • The interaction between hRad17 and these proteins exhibits characteristics similar to the RFC-PCNA interaction.
  • DNA damage was found to modulate the association of hRad17 with hRad1, hHus1, and hRad9.

Conclusions:

  • Provides the first experimental evidence for hRad17 interaction with PCNA-like proteins hRad1, hHus1, and hRad9.
  • Suggests a functional analogy between the hRad17-hRad1/hHus1/hRad9 complex and the RFC-PCNA clamp-loader system.
  • Highlights the role of this interaction in the DNA damage response pathway.

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