Regulation of DNA replication and repair proteins through interaction with the front side of proliferating cell

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Z O Jónsson1, R Hindges, U Hübscher

  • 1Department of Veterinary Biochemistry, University Zürich-Irchel, Winterthurerstrasse 190, CH-8057 Zürich, Switzerland.

The EMBO Journal
|May 26, 1998
PubMed

Insights

Proliferating cell nuclear antigen (PCNA) interacts with many proteins crucial for DNA replication. Mutating PCNA surface loops identified a key interaction site involved in binding multiple replication factors and suggested PCNA

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Genetics

Background:

  • Proliferating cell nuclear antigen (PCNA) is a key accessory factor for DNA polymerases.
  • PCNA interacts with a growing number of proteins involved in DNA replication and repair.

Purpose of the Study:

  • To characterize the interaction sites and functions of PCNA with various proteins.
  • To identify specific regions on PCNA responsible for binding to DNA polymerase delta, replication factor C, Fen1, p21, and DNA ligase I.

Main Methods:

  • Construction and analysis of four human PCNA mutants.
  • Targeting surface loops of the PCNA trimer by changing residues to alanine.
  • Assaying protein-protein interactions and functional consequences.

Main Results:

  • A specific region on the PCNA trimer surface, including the domain-connecting loop and loops near the C-termini, is critical for binding multiple replication proteins.
  • PCNA interaction with DNA ligase I inhibits DNA ligation.
  • DNA ligase I and Fen1 can inhibit DNA polymerase delta/PCNA-mediated DNA synthesis.
  • PCNA orientation at a 5' flap is necessary to stimulate Fen1 activity, suggesting C-termini face the direction of DNA synthesis.

Conclusions:

  • The identified PCNA surface region is a multifunctional interaction hub for DNA replication and repair proteins.
  • PCNA's orientation during replication is crucial for coordinating the activities of different enzymes.
  • These findings provide insights into the mechanism of DNA replication and the role of PCNA in coordinating enzymatic activities.

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