El volumen de ventas de los últimos tres años se encuentra:

This work describes the first detailed structural mechanism of Y family DNA polymerase nucleotide selection during different DNA replication contexts. The finger domain is directly involved in nucleotide selection by rejecting incoming NTP‟s and influencing hydrogen bonding and base staking potentials of incoming dNTP‟s. In addition to nucleotide selection, the finger domain allows the accommodation of bulky DNA adducts

via the solvent exposed major grove, facilitating lesion bypass. These results shed light on how Y family polymerase activity promotes cellular tolerance to DNA damaging agents and its contribution to carcinogenicity, aging, and evolution. Furthermore, our structural insights into Y family polymerase activity will aid in the design of novel drugs to inhibit or functionally alter these potentially therapeutic targets.

6.5

References

Albertella, M.R., Green, C.M., Lehmann, A.R. and O'Connor, M.J. (2005) A role for polymerase eta in the cellular tolerance to cisplatin-induced damage. Cancer Res,

65, 9799-9806.

Alt, A., Lammens, K., Chiocchini, C., Lammens, A., Pieck, J.C., Kuch, D., Hopfner, K.P. and Carell, T. (2007) Bypass of DNA lesions generated during anticancer treatment with cisplatin by DNA polymerase eta. Science, 318, 967-970.

Bauer, J., Xing, G., Yagi, H., Sayer, J.M., Jerina, D.M. and Ling, H. (2007) A structural gap in Dpo4 supports mutagenic bypass of a major benzo[a]pyrene dG adduct in DNA through template misalignment. Proc Natl Acad Sci U S A, 104, 14905- 14910.

Bienko, M., Green, C.M., Sabbioneda, S., Crosetto, N., Matic, I., Hibbert, R.G., Begovic, T., Niimi, A., Mann, M., Lehmann, A.R. and Dikic, I. (2010) Regulation of translesion synthesis DNA polymerase eta by monoubiquitination. Mol Cell, 37, 396-407.

Boudsocq, F., Kokoska, R.J., Plosky, B.S., Vaisman, A., Ling, H., Kunkel, T.A., Yang, W. and Woodgate, R. (2004) Investigating the role of the little finger domain of Y-family DNA polymerases in low fidelity synthesis and translesion replication. J Biol Chem, 279, 32932-32940.

Faili, A., Aoufouchi, S., Flatter, E., Gueranger, Q., Reynaud, C.A. and Weill, J.C. (2002) Induction of somatic hypermutation in immunoglobulin genes is dependent on DNA polymerase iota. Nature, 419, 944-947.

Hsu, G.W., Ober, M., Carell, T. and Beese, L.S. (2004) Error-prone replication of oxidatively damaged DNA by a high-fidelity DNA polymerase. Nature, 431, 217- 221.

Kim, S.H. and Michael, W.M. (2008) Regulated proteolysis of DNA polymerase eta during the DNA-damage response in C. elegans. Mol Cell, 32, 757-766.

Ling, H., Boudsocq, F., Woodgate, R. and Yang, W. (2001) Crystal structure of a Y- family DNA polymerase in action: a mechanism for error-prone and lesion-bypass replication. Cell, 107, 91-102.

Ling, H., Sayer, J.M., Plosky, B.S., Yagi, H., Boudsocq, F., Woodgate, R., Jerina, D.M. and Yang, W. (2004) Crystal structure of a benzo[a]pyrene diol epoxide adduct in a ternary complex with a DNA polymerase. Proc Natl Acad Sci U S A, 101, 2265- 2269.

Matsuda, T., Bebenek, K., Masutani, C., Hanaoka, F. and Kunkel, T.A. (2000) Low fidelity DNA synthesis by human DNA polymerase-eta. Nature, 404, 1011-1013. McCulloch, S.D., Kokoska, R.J., Masutani, C., Iwai, S., Hanaoka, F. and Kunkel, T.A.

(2004) Preferential cis-syn thymine dimer bypass by DNA polymerase eta occurs with biased fidelity. Nature, 428, 97-100.

Northam, M.R., Robinson, H.A., Kochenova, O.V. and Shcherbakova, P.V. (2009) Participation of DNA polymerase zeta in replication of undamaged DNA in Saccharomyces cerevisiae. Genetics, 184, 27-42.

Petta, T.B., Nakajima, S., Zlatanou, A., Despras, E., Couve-Privat, S., Ishchenko, A., Sarasin, A., Yasui, A. and Kannouche, P. (2008) Human DNA polymerase iota protects cells against oxidative stress. Embo J, 27, 2883-2895.

Prasad, R., Bebenek, K., Hou, E., Shock, D.D., Beard, W.A., Woodgate, R., Kunkel, T.A. and Wilson, S.H. (2003) Localization of the deoxyribose phosphate lyase active site in human DNA polymerase iota by controlled proteolysis. J Biol Chem, 278, 29649-29654.

Rechkoblit, O., Malinina, L., Cheng, Y., Geacintov, N.E., Broyde, S. and Patel, D.J. (2009) Impact of conformational heterogeneity of OxoG lesions and their pairing partners on bypass fidelity by Y family polymerases. Structure, 17, 725-736. Rechkoblit, O., Malinina, L., Cheng, Y., Kuryavyi, V., Broyde, S., Geacintov, N.E. and

Patel, D.J. (2006) Stepwise translocation of Dpo4 polymerase during error-free bypass of an oxoG lesion. PLoS Biol, 4, e11.

Sabouri, N., Viberg, J., Goyal, D.K., Johansson, E. and Chabes, A. (2008) Evidence for lesion bypass by yeast replicative DNA polymerases during DNA damage.

Nucleic Acids Res, 36, 5660-5667.

Shimizu, T., Azuma, T., Ishiguro, M., Kanjo, N., Yamada, S. and Ohmori, H. (2005) Normal immunoglobulin gene somatic hypermutation in Pol kappa-Pol iota double-deficient mice. Immunol Lett, 98, 259-264.

Terai, K., Abbas, T., Jazaeri, A.A. and Dutta, A. (2010) CRL4(Cdt2) E3 ubiquitin ligase monoubiquitinates PCNA to promote translesion DNA synthesis. Mol Cell, 37, 143-149.

Tomkinson, A.E., Chen, L., Dong, Z., Leppard, J.B., Levin, D.S., Mackey, Z.B. and Motycka, T.A. (2001) Completion of base excision repair by mammalian DNA ligases. Prog Nucleic Acid Res Mol Biol, 68, 151-164.

Vasilescu, J. and Figeys, D. (2006) Mapping protein-protein interactions by mass spectrometry. Curr Opin Biotechnol, 17, 394-399.

Watt, D.L., Utzat, C.D., Hilario, P. and Basu, A.K. (2007) Mutagenicity of the 1- nitropyrene-DNA adduct N-(deoxyguanosin-8-yl)-1-aminopyrene in mammalian cells. Chem Res Toxicol, 20, 1658-1664.

Wong, J.H., Fiala, K.A., Suo, Z. and Ling, H. (2008) Snapshots of a Y-family DNA polymerase in replication: substrate-induced conformational transitions and implications for fidelity of Dpo4. J Mol Biol, 379, 317-330.

Zeng, X., Winter, D.B., Kasmer, C., Kraemer, K.H., Lehmann, A.R. and Gearhart, P.J. (2001) DNA polymerase eta is an A-T mutator in somatic hypermutation of immunoglobulin variable genes. Nat Immunol, 2, 537-541.

Zhang, Y., Yuan, F., Wu, X., Wang, M., Rechkoblit, O., Taylor, J.S., Geacintov, N.E. and Wang, Z. (2000) Error-free and error-prone lesion bypass by human DNA polymerase kappa in vitro. Nucleic Acids Res, 28, 4138-4146.

Appendices

Appendix 1:

Curriculum Vitae

Name: Kevin Kirouac

Post-secondary University of Western Ontario

Education and London, Ontario, Canada

Degrees: 2001-2005 B.M.Sc Honors Biochemistry

The University of Western Ontario London, Ontario, Canada

2005-2010 Ph.D. Biochemsitry

Honours and Margaret P. Moffat Research Award

Awards: University of Western Ontario

2009

Schulich Graduate Scholarship University of Western Ontario 2007-2010

Graduate Research Scholarship University of Western Ontario 2005-2007

Publications:

Kirouac K, Ling H. (2009) Structural basis of error-prone replication and

stalling at a thymine base by Human DNA polymerase ι. EMBO J. 28: 1644-54 Xing G, Kirouac K, Shin YJ, Bell SD, Ling H. (2009) Structural insight into

recruitment of translesion DNA polymerase Dpo4 to sliding clamp PCNA. Mol Microbiol. 71: 678-91

Selected conference presentations:

“Error-free replication opposite an oxidative DNA lesion by a human Y family DNA polymerase.” American Crystallographic Association, July 25-30 2010, Chicago, Illinios, USA. Oral presentation

“Structural insights into error-prone replication and stalling opposite a template thymine base by a human DNA polymerase.” American Crystallographic Association, July 25-30 2009, Toronto, Canada. Oral presentation

“Structural insights into T template stalling and nucleotide mis-incorporation by human DNA polymerase iota.” 12th Annual DNA Replication and Repair Symposium, June 28 2008, Buffalo, Michigan, U.S.A. Oral presentation.