Requirement for PCNA and RPA in interstrand crosslink-induced DNA synthesis | Zendy

Lian Li | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Requirement for PCNA and RPA in interstrand crosslink-induced DNA synthesis

Author(s) -

Lian Li

Publication year - 2000

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/28.6.1424

Subject(s) - proliferating cell nuclear antigen , biology , dna replication , microbiology and biotechnology , dna , replication protein a , plasmid , replication factor c , in vitro , nucleotide , dna repair , dna synthesis , biochemistry , eukaryotic dna replication , dna binding protein , gene , transcription factor

Proliferating nuclear cell antigen (PCNA) and replication protein A (RPA) have proven to be essential elements in many aspects of DNA metabolism including replication, repair and recombination. We have developed an in vitro assay in which the presence of an interstrand crosslink stimulates the incorporation of radiolabeled nucleotides into both damaged and undamaged plasmid DNAs. Using this assay we have investigated the roles of PCNA and RPA in crosslink-induced DNA synthesis. p21, a potent inhibitor of PCNA, was found to strongly inhibit crosslink-induced incorporation. Addition of exogenous PCNA partially restored the resynthesis activity. Likewise, neutralization of RPA by monoclonal antibodies also inhibited incorporation, but the effect was somewhat more pronounced on the undamaged plasmid than the damaged plasmid. Addition of excess RPA also partially reversed antibody inhibition. These results indicate that both PCNA and RPA are required for efficient in vitro DNA resynthesis induced by interstrand crosslinks.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research