DNA polymerase eta, the product of the xeroderma pigmentosum variant gene and a target of p53, modulates the DNA damage checkpoint and p53 activation.

Abstract

DNA polymerase eta (PolH) is the product of the xeroderma pigmentosum variant (XPV) gene and a well-characterized Y-family DNA polymerase for translesion synthesis. Cells derived from XPV patients are unable to faithfully bypass UV photoproducts and DNA adducts and thus acquire genetic mutations. Here, we found that PolH can be up-regulated by DNA breaks induced by ionizing radiation or chemotherapeutic agents, and knockdown of PolH gives cells resistance to apoptosis induced by DNA breaks in multiple cell lines and cell types in a p53-dependent manner. To explore the underlying mechanism, we examined p53 activation upon DNA breaks and found that p53 activation is impaired in PolH knockdown cells and PolH-null primary fibroblasts. Importantly, reconstitution of PolH into PolH knockdown cells restores p53 activation. Moreover, we provide evidence that, upon DNA breaks, PolH is partially colocalized with phosphorylated ATM at gamma-H2AX foci and knockdown of PolH impairs ATM to phosphorylate Chk2 and p53. However, upon DNA damage by UV, PolH knockdown cells exhibit two opposing temporal responses: at the early stage, knockdown of PolH suppresses p53 activation and gives cells resistance to UV-induced apoptosis in a p53-dependent manner; at the late stage, knockdown of PolH suppresses DNA repair, leading to sustained activation of p53 and increased susceptibility to apoptosis in both a p53-dependent and a p53-independent manner. Taken together, we found that PolH has a novel role in the DNA damage checkpoint and that a p53 target can modulate the DNA damage response and subsequently regulate p53 activation.

• Authors: Gang Liu, Xinbin Chen
• Journal: Molecular and cellular biology (Mol Cell Biol) Vol. 26 Issue 4 Pg. 1398-413 (Feb 2006) ISSN: 0270-7306 [Print] United States
• PMID: 16449651 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Cell Cycle Proteins
• DNA, Complementary
• DNA-Binding Proteins
• RNA, Small Interfering
• TP53 protein, human
• Tumor Suppressor Protein p53
• Tumor Suppressor Proteins
• ATM protein, human
• Ataxia Telangiectasia Mutated Proteins
• Protein Serine-Threonine Kinases
• DNA-Directed DNA Polymerase
• Rad30 protein
• Camptothecin

Topics

• Apoptosis (drug effects, radiation effects)
• Ataxia Telangiectasia Mutated Proteins
• Base Sequence
• Camptothecin (pharmacology)
• Cell Cycle Proteins (metabolism)
• Cell Line
• DNA Damage
• DNA Repair
• DNA, Complementary (genetics)
• DNA-Binding Proteins (metabolism)
• DNA-Directed DNA Polymerase (genetics, metabolism)
• Genetic Variation
• Humans
• Models, Biological
• Protein Serine-Threonine Kinases (metabolism)
• RNA, Small Interfering (genetics)
• Tumor Suppressor Protein p53 (metabolism)
• Tumor Suppressor Proteins (metabolism)
• Ultraviolet Rays
• Xeroderma Pigmentosum (genetics, metabolism)