Dr. Thomas Melendy Microbiology and Immunology Associate Professor of Microbiology & Biochemistry 213 Biomedical Research Building Buffalo NY, 14214 Phone: (716) 829-3381 Email: tmelendy@buffalo.edu View map

DESCRIPTION OF INTERESTS

-The research interests of my laboratory are focused on the mechanism of human DNA replication and how DNA replication is controlled. We use model viral DNA replication systems, such as simian virus 40 (SV40) and papillomaviruses, both of which can replicate their DNA in human cells, as well as biochemical studies of human cellular proteins. Our studies on the mechanisms and regulation of DNA replication are intimately linked to our studies on novel papillomavirus and cellular DNA replication factors as well as our studies on how DNA damage inhibits DNA replication. Our studies on papillomavirus DNA replication are focused on identifying cellular proteins that papillomaviruses require to replicate their DNA. I have previously shown that all the cellular factors required for in vitro SV40 DNA replication are also required for papillomavirus DNA replication in vitro. However, they are not sufficient; additional cellular factors are required to support papillomavirus DNA replication. We are currently using a reconstitution assay to begin to purify those factors from human cells. In addition, a complementation assay, first developed in Dr. Louise Chow's laboratory at the University of Alabama at Birmingham, is currently being used to identify cellular factors in human 293 cell extracts that can complement human HeLa cell extracts for in vitro papillomavirus DNA replication. One of the ways in which we are studying the mechanism of DNA replication is by looking at the protein-protein interactions that take place between DNA replication factors at the replication fork. I previously demonstrated that an interaction between SV40-T -antigen (the viral DNA replication protein) and the human Replication Protein A complex appears to be important for priming during DNA synthesis. We are now studying the interactions between the papillomavirus DNA replication proteins and known cellular replication factors. Future plans include evaluating the biological significance of these interactions. Another search for novel human DNA replication factors entails the identification of human factors that are involved in the priming stages of DNA replication. We have utilized a novel priming inactivation assay to identify such an activity and are currently purifying this new DNA replication factor. Identification of this factor is very important in that this factor is apparently not required for current in vitro DNA replication systems which are based on SV40 and papillomaviruses. We have recently entered into a collaboration with Dr. Terry Beerman's laboratory at Roswell Park Cancer Institute to study the molecular effects of anticancer drugs on DNA replication. Dr. Beerman's laboratory has recently shown that a certain class of new chemotherapeutic anticancer drugs acts to block cellular DNA replication at very low levels of drug. This response seems to be similar to the very potent DNA damage dependent DNA replication arrest seen with ionizing radiation. We are currently investigating the mechanism by which these drugs inhibit DNA replication. In addition to elucidating how these anticancer drugs act to arrest cancer cell growth, we believe these studies will provide general information about how cells arrest DNA replication in response to DNA damage.

EDUCATION

1988	Doctor of Philosophy, Molecular Biology University of California at Los Angeles
1983	Bachelor of Science, Biological Sciences (Microbiology) Cornell University

PUBLICATIONS

Tezal M, Sullivan MA, Stoler D, Hyland A, Melendy T, Smaldino PJ, Rigual N, Loree TR; Chronic Periodontitis-Human Papillomavirus Synergy in Base of Tongue Cancers; Arch Otolaryngol Head Neck Surg; 2009 Apr; 135(4); 391-396

Tezal M, Sullivan MA, Stoler D, Hyland A, Melendy TE, Smaldino PJ, Rigual N, Loree T; Chronic Periodontitis, Smoking and Human Papillomavirus 16 Interaction in Base of Tongue Cancers; Arch Otolaryngol Head Neck Surg; 2008;

Liu, J.-S., Kuo, S.-K., Beerman, T.A., Melendy T; Induction of DNA damage responses by adozelesin is S phase-specific and dependent on active replication forks; Molec. Cancer Therapeutics; 2003 Jan; 2; 41-47

Melendy T, Li R; Chromatin remodeling and initiation of DNA replication.; Front Biosci; 2001 Sep; 6; 1048-1053

McHugh MM, Yin X, Kuo SR, Liu JS, Melendy T, Beerman TA; The cellular response to DNA damage induced by the enediynes C-1027 and neocarzinostatin includes hyperphosphorylation and increased nuclear retention of replication protein a (RPA) and trans inhibition of DNA replication.; Biochemistry; 2001 Apr; 40(15); 4792-4799

Dziegielewski J, Melendy T, Beerman TA; Bleomycin-induced alterations in DNA replication: relationship to DNA damage.; Biochemistry; 2001 Jan; 40(3); 704-711

J.S. Liu, S.R. Kuo, X. Yin, T. A. Beerman, Thomas Melendy; Damage by the Enediyne, C-1027, Results in the Inhibition of DNA Replication by Loss of Replication Protein a Function and Activation of DNA-Dependent Protein Kinase; Biochemistry; 2001 Jan; 40; 14661-14667

Loo YM, Melendy T; The majority of human replication protein A remains complexed throughout the cell cycle.; Nucleic Acids Res; 2000 Sep; 28(17); 3354-3360

Liu JS, Kuo SR, McHugh MM, Beerman TA, Melendy T; Adozelesin triggers DNA damage response pathways and arrests SV40 DNA replication through replication protein A inactivation.; J Biol Chem; 2000 Jan; 275(2); 1391-1397

McHugh MM, Kuo SR, Walsh-O'Beirne MH, Liu JS, Melendy T, Beerman TA; Bizelesin, a bifunctional cyclopropylpyrroloindole alkylating agent, inhibits simian virus 40 replication in trans by induction of an inhibitor.; Biochemistry; 1999 Aug; 38(35); 11508-11515

Dimitrova DS, Todorov IT, Melendy T, Gilbert DM; Mcm2, but not RPA, is a component of the mammalian early G1-phase prereplication complex.; J Cell Biol; 1999 Aug; 146(4); 709-722

Han Y, Loo YM, Militello KT, Melendy T; Interactions of the papovavirus DNA replication initiator proteins, bovine papillomavirus type 1 E1 and simian virus 40 large T antigen, with human replication protein A.; J Virol; 1999 Jun; 73(6); 4899-4907

Ni TH, McDonald WF, Zolotukhin I, Melendy T, Waga S, Stillman B, Muzyczka N; Cellular proteins required for adeno-associated virus DNA replication in the absence of adenovirus coinfection.; J Virol; 1998 Apr; 72(4); 2777-2787

Simmons DT, Melendy T, Usher D, Stillman B; Simian virus 40 large T antigen binds to topoisomerase I.; Virology; 1996 Aug; 222(2); 365-374

Melendy T, Sedman J, Stenlund A; Cellular factors required for papillomavirus DNA replication.; J Virol; 1995 Dec; 69(12); 7857-7867

Wun-Kim K, Upson R, Young W, Melendy T, Stillman B, Simmons DT; The DNA-binding domain of simian virus 40 tumor antigen has multiple functions.; J Virol; 1993 Dec; 67(12); 7608-7611

Brown GW, Melendy T, Ray DS; Replication protein A from the trypanosomatid Crithidia fasciculata is inactive in the primosome assembly step of SV40 DNA replication.; Mol Biochem Parasitol; 1993 Jun; 59(2); 323-325

Melendy T, Stillman B; An interaction between replication protein A and SV40 T antigen appears essential for primosome assembly during SV40 DNA replication.; J Biol Chem; 1993 Feb; 268(5); 3389-3395

Brown GW, Melendy TE, Ray DS; Conservation of structure and function of DNA replication protein A in the trypanosomatid Crithidia fasciculata.; Proc Natl Acad Sci U S A; 1992 Nov; 89(21); 10227-10231

Melendy T, Stillman B; Purification of DNA polymerase delta as an essential simian virus 40 DNA replication factor.; J Biol Chem; 1991 Jan; 266(3); 1942-1949

Tsurimoto T, Melendy T, Stillman B; Sequential initiation of lagging and leading strand synthesis by two different polymerase complexes at the SV40 DNA replication origin.; Nature; 1990 Aug; 346(6284); 534-539

Melendy T, Ray DS; Novobiocin affinity purification of a mitochondrial type II topoisomerase from the trypanosomatid Crithidia fasciculata.; J Biol Chem; 1989 Jan; 264(3); 1870-1876

Sheline C, Melendy T, Ray DS; Replication of DNA minicircles in kinetoplasts isolated from Crithidia fasciculata: structure of nascent minicircles.; Mol Cell Biol; 1989 Jan; 9(1); 169-176

Melendy T, Sheline C, Ray DS; Localization of a type II DNA topoisomerase to two sites at the periphery of the kinetoplast DNA of Crithidia fasciculata.; Cell; 1988 Dec; 55(6); 1083-1088

Melendy T, Ray DS; Purification and nuclear localization of a type I topoisomerase from Crithidia fasciculata.; Mol Biochem Parasitol; 1987 Jun; 24(2); 215-225

GRANTS

July 2001 to June 2002 UB Graduate Group Award Buffalo DNA Replication Graduate Group Thomas Melendy, William Ruyechan, Noreen Williams $3,000

January 2001 to December 2005 Mechanisms of DNA damage-triggered S phase checkpoints National Institutes of Health Thomas Melendy $169,000

April 2000 to May 2003 DNA replication arrest and DNA damage responses induced by AMGBs US Army Breat Cancer Initiative Thomas Melendy, Shu-Ru Kuo $143,855

November 1999 to October 2001 HPV DNA replication inhibitors Roche Discovery Welwyn Thomas Melendy $48,000

September 1999 to August 2004 HPV DNA replication: novel host factors National Institutes of Health Thomas Melendy $482,129

September 1999 to August 2004 Training in microbial pathogenesis National Institutes of Health Thomas Melendy, John Hay $162,556

May 1999 to June 2000 Effects of alkylating minor groove binders on DNA replication UB Multidisciplinary Pilot Project Program Thomas Melendy, Liu, Jen-Sing, Beerman, Terry $19,867

April 1998 to March 1999 The relationship between cellular responses to DNA damaging antitumor agents and inhibition of DNA replication Thomas Melendy, Beerman, Terry $148,000

January 1998 to December 2001 Cellular initiation proteins in eukaryotic DNA replication American Cancer Society Thomas Melendy $43,013

August 1997 to July 2002 Papillomavirus DNA replication: host cell factors National Institutes of Health Thomas Melendy $74,770

May 1997 to August 1998 Cell-cycle localization and association of the human Replication Protein A subunits UB Multidisciplinary Pilot Project Program Thomas Melendy, Berezney, Ronald $20,000

May 1995 to June 1996 Role of viral glycoproteins in infectious entry Thomas Melendy, Flanagan, Thomas $19,235