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Published on: December 30, 2016
Crystal structure of human PCNA in complex with the PIP box of DVC1
Yong Wang1, Min Xu2, Tao Jiang2
1National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, 19A Yuquan Road, Shijingshan District, Beijing 100049, China.
Abstract:
In higher eukaryotes, DVC1 (SPRTN, Spartan or C1orf124) is implicated in the translesion synthesis (TLS) pathway. DVC1 localizes to sites of DNA damage, binds to the proliferating cell nuclear antigen (PCNA) via its conserved PCNA-interacting motif (PIP box), and associates with ubiquitin selective segregase p97 and other factors, thus regulating translesion synthesis polymerases. Here, we report the crystal structure of human PCNA in complex with a peptide ((321)SNSHQNVLSNYFPRVS(336)) derived from human DVC1 that contains a unique YF type PIP box. Structural analysis reveals the detailed PIP box-PCNA interaction. Interestingly, substitution of Y331 with Phe severely reduces its PCNA binding affinity. These findings offer new insights into the determinants of PIP box for PCNA binding.
Insights
The study reveals how DVC1 protein binds to proliferating cell nuclear antigen (PCNA) through its unique PIP box. This interaction is crucial for DNA repair pathways and understanding protein-DNA binding mechanisms.
Area of Science:
- Molecular Biology
- Structural Biology
- Biochemistry
Background:
- DVC1 (SPRTN) is involved in the translesion synthesis (TLS) DNA repair pathway in eukaryotes.
- DVC1 interacts with proliferating cell nuclear antigen (PCNA) via a PCNA-interacting motif (PIP box).
Purpose of the Study:
- To determine the crystal structure of human PCNA in complex with a DVC1-derived peptide containing a YF type PIP box.
- To elucidate the detailed interaction between the PIP box of DVC1 and PCNA.
Main Methods:
- X-ray crystallography was used to obtain the structure of the human PCNA-DVC1 peptide complex.
- Site-directed mutagenesis was employed to substitute a key tyrosine residue (Y331) with phenylalanine.
Main Results:
- The crystal structure revealed the specific interactions between the unique YF type PIP box of DVC1 and human PCNA.
- Substitution of tyrosine at position 331 (Y331) with phenylalanine significantly reduced PCNA binding affinity.
Conclusions:
- The findings provide detailed insights into the structural determinants governing PIP box-PCNA binding.
- This structural information is critical for understanding DVC1's role in DNA repair and TLS regulation.
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