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Thomas Spratt
Title Associate Professor
Institution College of Medicine
Department Biochemistry and Molecular Biology
Division Biochemistry/Molecular Biology
Address 500 University Drive Hershey PA 17033
Mailbox: H171
Telephone 7175314623
Email
Background
Titles/roles

Associate Professor of Biochemistry and Molecular Biology
Director, IBIOS-Chemical Biology Option
Director, Biochemistry and Molecular Biology program
Member, PSU Cancer Institute
Member, PSU Center for Medical Genomics

Graduate Program Affliations:

Biomedical Sciences Graduate Program
Biochemistry and Molecular Genetics Option
Molecular Toxicology

Educational background:

BA, University of Rochester, 1978
Ph.D., University of Chicago, 1985
post-doc Ohio State University, 1985-86
post-doc, American Health Foundation. 1986-88

Narrative:

The Enzymology of DNA Repair and Mutagenesis

My research is based upon the belief that if we understand the mechanisms of cellular processes involved in mutagenesis and carcinogenesis we can design strategies for the prevention or treatment of cancer. My research is focused in projects elucidating the mechanisms of DNA repair, DNA replication, and mutagenesis.

My approach is to use organic synthesis and enzyme kinetics to examine the active site chemistry of the target enzymes. By this approach we are able to probe the structures of transitions states of enzymes reactions. By this approach, we expand upon the NMR and X-ray crystallography experiments, which can only examine ground state intermediates, to investigate the structures of the transition states.

A project typically consist of four phases: (1) formulation of a hypothesis based upon previous biochemical studies, especially structural studies using NMR or X-ray crystallography, (2) design of a substrate, typically an unnatural substrate, to test the hypothesis,. (3) synthesis of the substrate, and (4) kinetic evaluation of activity of substrate.

Mechanisms of fidelity and mutagenesis of DNA polymerases.

The long-term objective of this project is to understand the chemical interactions that govern the high fidelity replication of DNA. In the short-term, the goal of this research project is to test the hypothesis that specific interactions between DNA polymerases and the minor groove of DNA are crucial to the faithful replication of DNA.

The interactions between the polymerase and minor groove will be studied using a combination of amino acid substitution of the polymerase and atomic substitution of the DNA. Using atomic substitution of the DNA (3-deazaguanine and 1-(3-hydroxy-4-hydroxymethylcyclopentyl)uracil shown below) and site-directed mutagenesis of E. coli DNA polymerase I we have proposed that Arg668 makes a hydrogen bonding fork with the primer terminus and incoming dNTP. These interactions can serve as a sensor for Watson-Crick geometry at these positions.
Publications
1. Gowda AS, Krishnegowda G, Suo Z, Amin S, Spratt TE. Low fidelity bypass of O(2)-(3-pyridyl)-4-oxobutylthymine, the most persistent bulky adduct produced by the tobacco specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone by model DNA polymerases. Chem Res Toxicol. 2012 Jun 18; 25(6):1195-202.
  View in: PubMed
 
2. Jones SA, Boregowda R, Spratt TE, Hu J. In vitro epsilon RNA-dependent protein priming activity of human hepatitis B virus polymerase. J Virol. 2012 May; 86(9):5134-50.
  View in: PubMed
 
3. Jasti VP, Spratt TE, Basu AK. Tobacco-specific nitrosamine-derived O2-alkylthymidines are potent mutagenic lesions in SOS-induced Escherichia coli. Chem Res Toxicol. 2011 Nov 21; 24(11):1833-5.
  View in: PubMed
 
4. Olson KC, Sun D, Chen G, Sharma AK, Amin S, Ropson IJ, Spratt TE, Lazarus P. Characterization of dibenzo[a,l]pyrene-trans-11,12-diol (dibenzo[def,p]chrysene) glucuronidation by UDP-glucuronosyltransferases. Chem Res Toxicol. 2011 Sep 19; 24(9):1549-59.
  View in: PubMed
 
5. Krishnegowda G, Sharma AK, Krzeminski J, Gowda AS, Lin JM, Desai D, Spratt TE, Amin S. Facile syntheses of O(2)-[4-(3-pyridyl-4-oxobut-1-yl]thymidine, the major adduct formed by tobacco specific nitrosamine 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) in vivo, and its site-specifically adducted oligodeoxynucleotides. Chem Res Toxicol. 2011 Jun 20; 24(6):960-7.
  View in: PubMed
 
6. Sk UH, Prakasha Gowda AS, Crampsie MA, Yun JK, Spratt TE, Amin S, Sharma AK. Development of novel naphthalimide derivatives and their evaluation as potential melanoma therapeutics. Eur J Med Chem. 2011 Aug; 46(8):3331-8.
  View in: PubMed
 
7. Rubinson EH, Gowda AS, Spratt TE, Gold B, Eichman BF. An unprecedented nucleic acid capture mechanism for excision of DNA damage. Nature. 2010 Nov 18; 468(7322):406-11.
  View in: PubMed
 
8. Jones NR, Spratt TE, Berg AS, Muscat JE, Lazarus P, Gallagher CJ. Association studies of excision repair cross-complementation group 1 (ERCC1) haplotypes with lung and head and neck cancer risk in a Caucasian population. Cancer Epidemiol. 2011 Apr; 35(2):175-81.
  View in: PubMed
 
9. Zhong Q, Amin S, Lazarus P, Spratt TE. Differential repair of polycyclic aromatic hydrocarbon DNA adducts from an actively transcribed gene. DNA Repair (Amst). 2010 Sep 4; 9(9):1011-6.
  View in: PubMed
 
10. Prakasha Gowda AS, Polizzi JM, Eckert KA, Spratt TE. Incorporation of gemcitabine and cytarabine into DNA by DNA polymerase beta and ligase III/XRCC1. Biochemistry. 2010 Jun 15; 49(23):4833-40.
  View in: PubMed
 
11. Olson KC, Dellinger RW, Zhong Q, Sun D, Amin S, Spratt TE, Lazarus P. Functional characterization of low-prevalence missense polymorphisms in the UDP-glucuronosyltransferase 1A9 gene. Drug Metab Dispos. 2009 Oct; 37(10):1999-2007.
  View in: PubMed
 
12. Trostler M, Delier A, Beckman J, Urban M, Patro JN, Spratt TE, Beese LS, Kuchta RD. Discrimination between right and wrong purine dNTPs by DNA polymerase I from Bacillus stearothermophilus. Biochemistry. 2009 Jun 2; 48(21):4633-41.
  View in: PubMed
 
13. Cavanaugh NA, Urban M, Beckman J, Spratt TE, Kuchta RD. Identifying the features of purine dNTPs that allow accurate and efficient DNA replication by herpes simplex virus I DNA polymerase. Biochemistry. 2009 Apr 21; 48(15):3554-64.
  View in: PubMed
 
14. Chen KM, Sacks PG, Spratt TE, Lin JM, Boyiri T, Schwartz J, Richie JP, Calcagnotto A, Das A, Bortner J, Zhao Z, Amin S, Guttenplan J, El-Bayoumy K. Modulations of benzo[a]pyrene-induced DNA adduct, cyclin D1 and PCNA in oral tissue by 1,4-phenylenebis(methylene)selenocyanate. Biochem Biophys Res Commun. 2009 May 22; 383(1):151-5.
  View in: PubMed
 
15. DeCarlo L, Gowda AS, Suo Z, Spratt TE. Formation of purine-purine mispairs by Sulfolobus solfataricus DNA polymerase IV. Biochemistry. 2008 Aug 5; 47(31):8157-64.
  View in: PubMed
 
16. Chen G, Dellinger RW, Sun D, Spratt TE, Lazarus P. Glucuronidation of tobacco-specific nitrosamines by UGT2B10. Drug Metab Dispos. 2008 May; 36(5):824-30.
  View in: PubMed
 
17. Chen KM, Spratt TE, Stanley BA, De Cotiis DA, Bewley MC, Flanagan JM, Desai D, Das A, Fiala ES, Amin S, El-Bayoumy K. Inhibition of nuclear factor-kappaB DNA binding by organoselenocyanates through covalent modification of the p50 subunit. Cancer Res. 2007 Nov 1; 67(21):10475-83.
  View in: PubMed
 
18. Coulter R, Blandino M, Tomlinson JM, Pauly GT, Krajewska M, Moschel RC, Peterson LA, Pegg AE, Spratt TE. Differences in the rate of repair of O6-alkylguanines in different sequence contexts by O6-alkylguanine-DNA alkyltransferase. Chem Res Toxicol. 2007 Dec; 20(12):1966-71.
  View in: PubMed
 
19. Beckman J, Kincaid K, Hocek M, Spratt T, Engels J, Cosstick R, Kuchta RD. Human DNA polymerase alpha uses a combination of positive and negative selectivity to polymerize purine dNTPs with high fidelity. Biochemistry. 2007 Jan 16; 46(2):448-60.
  View in: PubMed
 
20. Kretulskie AM, Spratt TE. Structure of purine-purine mispairs during misincorporation and extension by Escherichia coli DNA polymerase I. Biochemistry. 2006 Mar 21; 45(11):3740-6.
  View in: PubMed
 
21. Wolfle WT, Washington MT, Kool ET, Spratt TE, Helquist SA, Prakash L, Prakash S. Evidence for a Watson-Crick hydrogen bonding requirement in DNA synthesis by human DNA polymerase kappa. Mol Cell Biol. 2005 Aug; 25(16):7137-43.
  View in: PubMed
 
22. McCain MD, Meyer AS, Schultz SS, Glekas A, Spratt TE. Fidelity of mispair formation and mispair extension is dependent on the interaction between the minor groove of the primer terminus and Arg668 of DNA polymerase I of Escherichia coli. Biochemistry. 2005 Apr 19; 44(15):5647-59.
  View in: PubMed
 
23. Meyer AS, Blandino M, Spratt TE. Escherichia coli DNA polymerase I (Klenow fragment) uses a hydrogen-bonding fork from Arg668 to the primer terminus and incoming deoxynucleotide triphosphate to catalyze DNA replication. J Biol Chem. 2004 Aug 6; 279(32):33043-6.
  View in: PubMed
 
24. Guttenplan JB, Spratt TE, Khmelnitsky M, Kosinska W, Desai D, El-Bayoumy K. Effects of 3H-1,2-dithiole-3-thione, 1,4-phenylenebis(methylene)selenocyanate, and selenium-enriched yeast individually and in combination on benzo[a]pyrene-induced mutagenesis in oral tissue and esophagus in lacZ mice. Mutat Res. 2004 Apr 11; 559(1-2):199-210.
  View in: PubMed
 
25. Meyer AS, McCain MD, Fang Q, Pegg AE, Spratt TE. O6-alkylguanine-DNA alkyltransferases repair O6-methylguanine in DNA with Michaelis-Menten-like kinetics. Chem Res Toxicol. 2003 Nov; 16(11):1405-9.
  View in: PubMed
 
26. Washington MT, Wolfle WT, Spratt TE, Prakash L, Prakash S. Yeast DNA polymerase eta makes functional contacts with the DNA minor groove only at the incoming nucleoside triphosphate. Proc Natl Acad Sci U S A. 2003 Apr 29; 100(9):5113-8.
  View in: PubMed
 
27. Spratt TE. Identification of hydrogen bonds between Escherichia coli DNA polymerase I (Klenow fragment) and the minor groove of DNA by amino acid substitution of the polymerase and atomic substitution of the DNA. Biochemistry. 2001 Mar 6; 40(9):2647-52.
  View in: PubMed
 
28. Spratt TE, Schultz SS, Levy DE, Chen D, Schlüter G, Williams GM. Different mechanisms for the photoinduced production of oxidative DNA damage by fluoroquinolones differing in photostability. Chem Res Toxicol. 1999 Sep; 12(9):809-15.
  View in: PubMed
 
29. Spratt TE, Wu JD, Levy DE, Kanugula S, Pegg AE. Reaction and binding of oligodeoxynucleotides containing analogues of O6-methylguanine with wild-type and mutant human O6-alkylguanine-DNA alkyltransferase. Biochemistry. 1999 May 25; 38(21):6801-6.
  View in: PubMed
 
30. Wang L, Spratt TE, Pegg AE, Peterson LA. Synthesis of DNA oligonucleotides containing site-specifically incorporated O6-[4-oxo-4-(3-pyridyl)butyl]guanine and their reaction with O6-alkylguanine-DNA alkyltransferase. Chem Res Toxicol. 1999 Feb; 12(2):127-31.
  View in: PubMed
 
31. Nunes MG, Desai D, Koehl W, Spratt TE, Guengerich FP, Amin S. Inhibition of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) metabolism in human hepatic microsomes by ipomeanol analogs--an exploratory study. Cancer Lett. 1998 Jul 17; 129(2):131-8.
  View in: PubMed
 
32. Spratt TE, Zydowsky TM, Floss HG. Stereochemistry of the in vitro and in vivo methylation of DNA by (R)- and (S)-N-[2H1,3H]methyl-N-nitrosourea and (R)- and (S)-N-nitroso-N-[2H1,3H]methyl-N-methylamine. Chem Res Toxicol. 1997 Dec; 10(12):1412-9.
  View in: PubMed
 
33. Spratt TE. Klenow fragment-DNA interaction required for the incorporation of nucleotides opposite guanine and O6-methylguanine. Biochemistry. 1997 Oct 28; 36(43):13292-7.
  View in: PubMed
 
34. Hecht SS, Spratt TE, Trushin N. Absolute configuration of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol formed metabolically from 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone. Carcinogenesis. 1997 Sep; 18(9):1851-4.
  View in: PubMed
 
35. Spratt TE, Levy DE. Structure of the hydrogen bonding complex of O6-methylguanine with cytosine and thymine during DNA replication. Nucleic Acids Res. 1997 Aug 15; 25(16):3354-61.
  View in: PubMed
 
36. Rosen JE, Chen D, Prahalad AK, Spratt TE, Schluter G, Williams GM. A fluoroquinolone antibiotic with a methoxy group at the 8 position yields reduced generation of 8-oxo-7,8-dihydro-2'-deoxyguanosine after ultraviolet-A irradiation. Toxicol Appl Pharmacol. 1997 Aug; 145(2):381-7.
  View in: PubMed
 
37. Wang L, Spratt TE, Liu XK, Hecht SS, Pegg AE, Peterson LA. Pyridyloxobutyl adduct O6-[4-oxo-4-(3-pyridyl)butyl]guanine is present in 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone-treated DNA and is a substrate for O6-alkylguanine-DNA alkyltransferase. Chem Res Toxicol. 1997 May; 10(5):562-7.
  View in: PubMed
 
38. Liu XK, Spratt TE, Murphy SE, Peterson LA. Pyridyloxobutylation of guanine residues by 4-[(acetoxymethyl)nitrosamino]-1-(3-pyridyl)-1-butanone generates substrates of O6-alkylguanine-DNA alkyltransferase. Chem Res Toxicol. 1996 Sep; 9(6):949-53.
  View in: PubMed
 
39. Chen W, Weisburger JH, Fiala ES, Spratt TE, Carmella SG, Chen D, Hecht SS. Gastric carcinogenesis: 2-chloro-4-methylthiobutanoic acid, a novel mutagen in salted, pickled Sanma hiraki fish, or similarly treated methionine. Chem Res Toxicol. 1996 Jan-Feb; 9(1):58-66.
  View in: PubMed
 
40. Chen W, Weisburger JH, Fiala ES, Carmella SG, Chen D, Spratt TE, Hecht SS. Unexpected mutagen in fish. Nature. 1995 Apr 13; 374(6523):599.
  View in: PubMed
 
41. Spratt TE, Campbell CR. Synthesis of oligodeoxynucleotides containing analogs of O6-methylguanine and reaction with O6-alkylguanine-DNA alkyltransferase. Biochemistry. 1994 Sep 20; 33(37):11364-71.
  View in: PubMed
 
42. Hecht SS, Peterson LA, Spratt TE. Tobacco-specific nitrosamines. IARC Sci Publ. 1994; (125):91-106.
  View in: PubMed
 
43. Yang YM, Rutberg SE, Luo FC, Spratt TE, Halaban R, Ferrone S, Ronai Z. A transcriptional inhibitor induced in human melanoma cells upon ultraviolet irradiation. Cell Growth Differ. 1993 Jul; 4(7):595-602.
  View in: PubMed
 
44. Spratt TE, de los Santos H. Reaction of O6-alkylguanine-DNA alkyltransferase with O6-methylguanine analogues: evidence that the oxygen of O6-methylguanine is protonated by the protein to effect methyl transfer. Biochemistry. 1992 Apr 14; 31(14):3688-94.
  View in: PubMed
 
45. Carmella SG, Kagan SS, Spratt TE, Hecht SS. Evaluation of cysteine adduct formation in rat hemoglobin by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and related compounds. Cancer Res. 1990 Sep 1; 50(17):5453-9.
  View in: PubMed
 
46. Spratt TE, Peterson LA, Confer WL, Hecht SS. Solvolysis of model compounds for alpha-hydroxylation of N'-nitrosonornicotine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone: evidence for a cyclic oxonium ion intermediate in the alkylation of nucleophiles. Chem Res Toxicol. 1990 Jul-Aug; 3(4):350-6.
  View in: PubMed
 
47. Luks HJ, Spratt TE, Vavrek MT, Roland SF, Weisburger JH. Identification of sulfate and glucuronic acid conjugates of the 5-hydroxy derivative as major metabolites of 2-amino-3-methylimidazo[4,5-f]quinoline in rats. Cancer Res. 1989 Aug 15; 49(16):4407-11.
  View in: PubMed
 
48. Spratt TE, Trushin N, Lin D, Hecht SS. Analysis for N2-(pyridyloxobutyl)deoxyguanosine adducts in DNA of tissues exposed to tritium-labeled 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-nitrosonornicotine. Chem Res Toxicol. 1989 May-Jun; 2(3):169-73.
  View in: PubMed
 
49. Hecht SS, Carmella SG, Trushin N, Spratt TE, Foiles PG, Hoffmann D. Approaches to the development of assays for interaction of tobacco-specific nitrosamines with haemoglobin and DNA. IARC Sci Publ. 1988; (89):121-8.
  View in: PubMed
 
50. Hecht SS, Spratt TE, Trushin N. Evidence for 4-(3-pyridyl)-4-oxobutylation of DNA in F344 rats treated with the tobacco-specific nitrosamines 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-nitrosonornicotine. Carcinogenesis. 1988 Jan; 9(1):161-5.
  View in: PubMed
 
 
Keyword
Last Name
Institution
    
 
 
 
Keywords   
Guanine
DNA Repair
DNA Polymerase I
DNA Damage
DNA
See all (246) keywords
Co-Authors  
Amin, Shantu
Desai, Dhimant
Gowda, Prakasha
Pegg, Anthony
Sharma, Arun
See all (24) people
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