Defects in Nucleotide Excision Repair (NER) have been linked directly to numerous human diseases including xeroderma pigmentosum, Cockayne syndrome and cancer. The mechanism of DNA damage recognition remains poorly understood in all cellular models. A central feature to the conundrum of damage recognition is whether localized distortion of the DNA duplex, possibly by the unpairing of base pairs near the damage site, is the crucial step enabling a site of DNA damage to be identified and subsequently removed by the NER machinery. If the unpairing of base pairs near the damage site is truly a crucial feature in identifying damage then it follows directly that sequences immediately surrounding any form of damage will bear on the damage recognizability, i.e. sequences that unpair more easily should be more easily recognized. The proposed study has two goals. The primary goal will be to determine in Saccharomyces cerevesiae if here is a DNA sequence context preference for recognition of two types of DNA damage known to be repaired exclusively by NER:2-acetylaminofluorene-DNA adducts and benzo[a]pyrene diol epoxide-DNA adducts. The recognizability of both forms of damage will be compared within a panel of DNA substrates that differ in the sequence context of the damage site. The secondary goal of this project is to make a correlation with the NER proteins themselves by identifying specifically which protein components within the NER machinery give rise to any observed sequence context preference.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32CA083314-02
Application #
6174091
Study Section
Special Emphasis Panel (ZRG1-SSS-1 (01))
Program Officer
Lohrey, Nancy
Project Start
2000-08-03
Project End
Budget Start
2000-08-03
Budget End
2001-08-02
Support Year
2
Fiscal Year
2000
Total Cost
$37,516
Indirect Cost
Name
University of Texas Sw Medical Center Dallas
Department
Pathology
Type
Schools of Medicine
DUNS #
City
Dallas
State
TX
Country
United States
Zip Code
75390
Fischhaber, Paula L; Gerlach, Valerie L; Feaver, William J et al. (2002) Human DNA polymerase kappa bypasses and extends beyond thymine glycols during translesion synthesis in vitro, preferentially incorporating correct nucleotides. J Biol Chem 277:37604-11
Gerlach, V L; Feaver, W J; Fischhaber, P L et al. (2001) Purification and characterization of pol kappa, a DNA polymerase encoded by the human DINB1 gene. J Biol Chem 276:92-8
Gerlach, V L; Feaver, W J; Fischhaber, P L et al. (2000) Human DNA polymerase kappa: a novel DNA polymerase of unknown biological function encoded by the DINB1 gene. Cold Spring Harb Symp Quant Biol 65:41-9