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Human DNA glycosylase NEIL1 but not NEIL2 preferentially collaborates with proteins of DNA replication to carry out LP‐BER in replicating cells
Author(s) -
Hegde Muralidhar L,
Hegde Pavana M,
Theriot Corey A,
Tomkinson Alan A,
Li GuMin,
Hazra Tapas K,
Mitra Sankar
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.876.4
Subject(s) - base excision repair , dna replication , proliferating cell nuclear antigen , biology , dna glycosylase , microbiology and biotechnology , dna repair , dna , dna damage , dna polymerase , genetics
We have previously shown that NEIL1, activated in S‐phase, functionally interacts with the PCNA, flap endonuclease 1 (FEN1), replication protein A (RPA), suggesting its preferential involvement in repairing replicating genome. NEIL1 along with NEIL2, has higher activity with damaged base in fork or bubble DNA that mimics replication/transcription intermediates. Here we show that both NEIL‐FLAG immunoprecipitates (IPs) from human cells contained replicative DNA polymeraseδ (Polδ), replication factor‐C (RFC) and DNA ligaseI (LigI), along with PCNA and FEN1. However, the NEIL1‐IP contained ~3–5 fold higher level of these proteins than the NEIL2 IP. Although, both the NEILs binarily interact with these proteins, only NEIL1 activates Polδ and RFC stimulates NEIL1 but not NEIL2. NEIL1 IP eluate could efficiently repair a base lesion via long‐patch repair (LP‐BER) pathway. We reconstituted NEIL1‐initiated LP‐BER using purified proteins and showed the dependence on PCNA and its loading by RF‐C. Repair initiated by C‐terminally truncated NEIL1 lacking the interaction domain was significantly less efficient compared to that initiated with full‐length NEIL1. Further, NEIL1 has significantly higher ability to repair 5‐OHU using replication proteins. These results showed that NEIL1 preferentially collaborates with proteins of DNA replication to carry out a distinct sub pathway of BER during DNA replication. (USPHS PO1 CA 92584 and APDA fellowship to MLH).