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L2DTL/CDT2 interacts with the CUL4/DDB1 complex and PCNA and regulates CDT1 proteolysis in response to DNA damage

Leigh Ann Higa1, Damon Banks, Min Wu

  • 1Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06520, USA.

Cell Cycle (Georgetown, Tex.)
|July 25, 2006
PubMed

Insights

The CUL4 ubiquitin E3 ligase complex

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Cullin 4 (CUL4) proteins are central to ubiquitin E3 ligase complexes.
  • These complexes regulate critical cellular processes including cell cycle, DNA replication, and DNA damage response.

Purpose of the Study:

  • To elucidate the composition of the CUL4 ubiquitin E3 ligase complex.
  • To identify novel interacting partners and their roles in DNA damage response.

Main Methods:

  • Anti-CUL4 antibody affinity chromatography followed by mass spectrometry.
  • Immuno-affinity chromatography using anti-L2DTL antibodies.
  • In vivo interaction studies and proteolysis assays.

Main Results:

  • Identified a novel WD40 domain protein, L2DTL (homologue of Drosophila L2DTL and fission yeast CDT2), interacting with CUL4 and DDB1.
  • L2DTL interacts with CDT1 and its loss suppresses CDT1 proteolysis post-DNA damage.
  • PCNA also interacts with CDT1, and its loss similarly suppresses CDT1 proteolysis.
  • L2DTL inactivation leads to DDB1 dissociation from the CUL4 complex.

Conclusions:

  • L2DTL and PCNA are key components of the CUL4/DDB1 complex involved in regulating CDT1 degradation.
  • This interaction is crucial for the cellular response to DNA damage.

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