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Poly (ADP‐ribose) polymerase (PARP) is essential for sulfur mustard‐induced DNA damage repair, but has no role in DNA ligase activation
Author(s) -
Bhat K. Ramachandra,
Benton Betty J.,
Ray Radharaman
Publication year - 2006
Publication title -
journal of applied toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.784
H-Index - 87
eISSN - 1099-1263
pISSN - 0260-437X
DOI - 10.1002/jat.1161
Subject(s) - poly adp ribose polymerase , dna ligase , dna repair , polymerase , dna damage , chemistry , sulfur mustard , dna , microbiology and biotechnology , dna polymerase mu , biochemistry , dna polymerase , biology , toxicity , circular bacterial chromosome , organic chemistry
Concurrent activation of poly (ADP‐ribose) polymerase (PARP) and DNA ligase was observed in cultured human epidermal keratinocytes (HEK) exposed to the DNA alkylating compound sulfur mustard (SM), suggesting that DNA ligase activation could be due to its modification by PARP. Using HEK, intracellular 3 H‐labeled NAD + ( 3 H‐adenine) was metabolically generated and then these cells were exposed to SM (1 m M ). DNA ligase I isolated from these cells was not 3 H‐labeled, indicating that DNA ligase I is not a substrate for (ADP‐ribosyl)ation by PARP. In HEK, when PARP was inhibited by 3‐amino benzamide (3‐AB, 2 m M ), SM‐activated DNA ligase had a half‐life that was four‐fold higher than that observed in the absence of 3‐AB. These results suggest that DNA repair requires PARP, and that DNA ligase remains activated until DNA damage repair is complete. The results show that in SM‐exposed HEK, DNA ligase I is activated by phosphorylation catalysed by DNA‐dependent protein kinase (DNA‐PK). Therefore, the role of PARP in DNA repair is other than that of DNA ligase I activation. By using the DNA ligase I phosphorylation assay and decreasing PARP chemically as well as by PARP anti‐sense mRNA expression in the cells, it was confirmed that PARP does not modify DNA ligase I. In conclusion, it is proposed that PARP is essential for efficient DNA repair; however, PARP participates in DNA repair by altering the chromosomal structure to make the DNA damage site(s) accessible to the repair enzymes. Published in 2006 by John Wiley & Sons, Ltd.