Accuracy of thymine-thymine dimer bypass by Saccharomyces cerevisiae DNA polymerase eta.

The Saccharomyces cerevisiae RAD30 gene functions in error-free replication of UV-damaged DNA. RAD30 encodes a DNA polymerase, Pol eta, which inserts two adenines opposite the two thymines of a cis-syn thymine-thymine (T-T) dimer. Here we use steady-state kinetics to determine the accuracy of DNA synthesis opposite the T-T dimer. Surprisingly, the accuracy of DNA synthesis opposite the damaged DNA is nearly indistinguishable from that opposite nondamaged DNA, with frequencies of misincorporation of about 10(-2) to 10(-3). These studies support the hypothesis that unlike most DNA polymerases, Pol eta is able to tolerate distortions in DNA resulting from damage, which then enables the polymerase to utilize the intrinsic base pairing ability of the T-T dimer.

[1]  Robert E. Johnson,et al.  hRAD30 mutations in the variant form of xeroderma pigmentosum. , 1999, Science.

[2]  Chikahide Masutani,et al.  The XPV (xeroderma pigmentosum variant) gene encodes human DNA polymerase η , 1999, Nature.

[3]  Robert E. Johnson,et al.  Requirement of DNA Polymerase Activity of Yeast Rad30 Protein for Its Biological Function* , 1999, The Journal of Biological Chemistry.

[4]  Robert E. Johnson,et al.  Efficient bypass of a thymine-thymine dimer by yeast DNA polymerase, Poleta. , 1999, Science.

[5]  E. G. Frank,et al.  Biochemical basis of SOS-induced mutagenesis in Escherichia coli: reconstitution of in vitro lesion bypass dependent on the UmuD'2C mutagenic complex and RecA protein. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  Z. Livneh,et al.  The mutagenesis proteins UmuD' and UmuC prevent lethal frameshifts while increasing base substitution mutations. , 1998, Molecular cell.

[7]  J. Vos,et al.  Defective bypass replication of a leading strand cyclobutane thymine dimer in xeroderma pigmentosum variant cell extracts. , 1998, Cancer Research.

[8]  D. Fygenson,et al.  DNA polymerase fidelity: from genetics toward a biochemical understanding. , 1998, Genetics.

[9]  W. Kaufmann,et al.  Replication Fork Bypass of a Pyrimidine Dimer Blocking Leading Strand DNA Synthesis* , 1997, The Journal of Biological Chemistry.

[10]  B. Edgar,et al.  Developmental Control of Cell Cycle Regulators: A Fly's Perspective , 1996, Science.

[11]  C. Lawrence,et al.  Thymine-Thymine Dimer Bypass by Yeast DNA Polymerase ζ , 1996, Science.

[12]  D. Patel,et al.  CONTRASTING STRUCTURAL IMPACTS INDUCED BY cis‐syn CYCLOBUTANE DIMER AND (6–4) ADDUCT IN DNA DUPLEX DECAMERS: IMPLICATION IN MUTAGENESIS AND REPAIR ACTIVITY , 1995, Photochemistry and photobiology.

[13]  P. V. Hippel,et al.  On the Processivity of Polymerases a , 1994, Annals of the New York Academy of Sciences.

[14]  H. L. Waters,et al.  Ultraviolet hypermutability of a shuttle vector propagated in xeroderma pigmentosum variant cells. , 1993, The Journal of investigative dermatology.

[15]  C. Lawrence,et al.  The frequency and accuracy of replication past a thymine-thymine cyclobutane dimer are very different in Saccharomyces cerevisiae and Escherichia coli , 1993, Journal of bacteriology.

[16]  J. Taylor,et al.  Site-specific effect of thymine dimer formation on dAn.dTn tract bending and its biological implications. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Mccormick,et al.  Xeroderma pigmentosum variant cells are less likely than normal cells to incorporate dAMP opposite photoproducts during replication of UV-irradiated plasmids. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[18]  B. Strauss.  The ‘A rule’ of mutagen specificity: A consequence of DNA polymerase bypass of non‐instructional lesions? , 1991, BioEssays : news and reviews in molecular, cellular and developmental biology.

[19]  G. A. van der Marel,et al.  1H NMR study of the exchangeable protons of the duplex d(GCGTTGCG).d(CGCAACGC) containing a thymine photodimer. , 1987, Nucleic acids research.

[20]  A. M. Pedrini,et al.  Determination of pyrimidine dimer unwinding angle by measurement of DNA electrophoretic mobility. , 1982, Journal of molecular biology.

[21]  A. Sancar.  DNA excision repair. , 1996, Annual review of biochemistry.

[22]  S. Creighton,et al.  Gel fidelity assay measuring nucleotide misinsertion, exonucleolytic proofreading, and lesion bypass efficiencies. , 1995, Methods in enzymology.

[23]  R. Bambara,et al.  [21] Methods of analyzing processivity , 1995 .

[24]  H. Echols,et al.  Fidelity mechanisms in DNA replication. , 1991, Annual review of biochemistry.

[25]  A. Fersht.  Enzyme structure and mechanism , 1977 .