Jeffrey DeStefano

Professor

Contact

Emailjdestefa@umd.edu

Office Phone: (301) 405-5449

Fax: (301) 314-1248

Office Address: 3130 Biosciences Research Building

URLhttp://www.clfs.umd.edu/cbmg/faculty/destefano/lab/index.htm

Graduate Program Affiliations

  • BISI - BISI-Molecular & Cellular Biology (MOCB)

Research Interests

HIV and poliovirus replication and recombination

Education

Ph.D.- University of Connecticut, 1990

Peer-reviewed Publications

1.     DeStefano, J., Olmstead, E., Panniers, R., and Lucas-Lenard, J. (1990) The Alpha Subunit of Eukaryotic Initiation Factor 2 is Phosphorylated in Mengovirus-Infected Mouse L Cells. J. Virol. 64, 4445-4453.

2.  DeStefano, J. J., Buiser, R.G., Mallaber, L. M., Myers, T. W., Bambara, R. A., and Fay, P. J. (1991) Polymerization and RNase H Activities of the Reverse Transcriptases from Avian Myeloblastosis, Human Immunodeficiency, and Moloney Murine Leukemia Viruses Are Functionally Uncoupled. J. Biol. Chem. 266, 7423-7431.

3.  Buiser, R. G., DeStefano, J. J., Mallaber, L. M., Fay, P. J., and Bambara, R. A. (1991) Requirements for the Catalysis of Strand Transfer Synthesis by Retroviral DNA Polymerases. J. Biol. Chem. 266,13103-13109.

4.  DeStefano, J. J., Buiser, R. G., Mallaber, L. M., Bambara, R. A., and Fay, P. J. (1991) Human Immunodeficiency Virus Reverse Transcriptase Possesses a 3' to 5' Endonuclease Activity. J.Biol. Chem. 266, 24295-24301.

5.  DeStefano, J. J., Buiser, R. G., Mallaber, L. M., Fay, P. J., and Bambara, R. A. (1992) Parameters that Influence Processive Synthesis and Site Specific Termination by Human Immunodeficiency Virus Reverse Transcriptase on RNA and DNA Tem plates. Biochim. Biophys. Acta.1131, 270-280.

6.  DeStefano, J. J., Mallaber, L. M., Rodriguez-Rodriguez, L., Fay, P. J., and Bambara, R. A. (1992) Requirments for Strand Transfer Between Internal Regions of Heteropolymer Templates by Human Immunodeficiency Virus Reverse Transcriptase. J. Virol. 66, 6370-6378.

7.  DeStefano, J. J., Bambara, R. A., and Fay, P. J. (1992) Parameters That Influence the Binding of Human Immunodeficiency Virus Reverse Transcriptase to Nucleic Acid Structures. Biochemistry 32, 6908-6915.

8.  DeStefano, J. J., Mallaber, L. M., Fay, P. J., and Bambara, R. A. (1993) Determinants of the RNase H Cleavage Specificity of Human Immunodeficiency Virus Reverse Transcriptase. Nuc. Acids Res.21,4330-4338.

9.  DeStefano, J. J., Bambara, P. J., and Fay, P. J. (1994) The Mechanism of Human Immunodeficiency Virus Reverse Transcriptase-catalyzed Strand Transfer Between Internal Regions of Heteropolymeric RNA Templates.J. Biol Chem. 269, 161-168.

10. DeStefano, J. J. (1994) Kinetic Analysis of the Catalysis of Strand Transfer from Internal Regions of Heteropolymeric RNA Templates by Human Immunodeficiency Virus Reverse Transcriptae. J. Mol. Biol.243, 558-567.

11. DeStefano, J. J., Wu, W., Seehra, J., McCoy, J., Laston, D., Albone, E., Fay, P.J. and Bambara, R.A. (1994) Characterization of an RNaseH Deficient Mutant of Human Immunodeficiency Virus-1 Reverse Transcriptase Having an Aspartate to Asparagine Change at Position 498. Biochem. Biophys. Acta., 1219, 380-388.

12. DeStefano, J. J., Mallaber, L. M., Fay, P. J., and Bambara, R. A. (1994) Quantitative Analysis of RNA Cleavage During RNA-Directed DNA Synthesis by Human Immunodeficiency and Avian Myeloblastosis Virus Reverse Transcriptases. Nuc. Acids. Res. 22, 3793-3800.

13. DeStefano, J. J. (1995) The orientation of binding of human immunodeficiency virus reverse transcriptase on nucleic acid hybrids.Nuc. Acids Res. 23, 3901-3908.

14. DeStefano, J. J. (1995) Human immunodeficiency virus nucleocapsid protein stimulates strand transfer from internal regions of heteropolymeric RNA. Arch. Virol. 140:1775-1789.

15. DeStefano, J. J. (1996) Interactions of human immunodeficiency virus nucleocapsid protein with a structure mimicking a replication intermediate. J. Biol. Chem. 271, 16350-16356.

16. Diaz, L., and DeStefano, J. J. (1996) Strand transfer is enhanced by mismatched nucleotides at the 3' primer terminus: A possible link between HIV-RT fidelity and recombination. Nuc. Acids Res. 24, 3086-3092.

17. DeStefano, J. J., Roberts, B., and Shriner, D. (1997) The mechanism of retroviral recombination: the role of sequences proximal to the point of strand transfer. Arch. Virol. 142, 1797-1812.

18. DeStefano, J., Ghosh, J., Prasad, B., and Raja, A. (1998) High fidelity of internal strand transfer catalyzed by human immunodeficiency virus reverse transcriptase. J. Biol Chem. 273, 1483-1489.

19. Raja, A., and DeStefano, J. J. (1999) Kinetic analysis of the effect of HIV nucleocapsid protein (NCp) on retroviral strand transfer reactions.Biochemistry 38, 5178-5184.

20. Diaz, L., Cristofaro, J. V., and DeStefano, J. J. (2000) Human immunodeficiency virus resverse transcriptase base misincorporations can promote strand transfer. Arch. Virol. 145, 1117-1131.

21. DeStefano, J. J. Raja, A., and Crisotfaro, J. V. (2000) In vitro strand transfer from broken RNAs results in mismatch but not frameshift mutations. Virology 276, 7-15.

22. Rodriguez-Wells, V., Plotch, S. J., and DeStefano, J. J. (2001) Determination of the mutation rate of poliovirus RNA-dependent RNA polymerase. Virus Res. 74, 119-1132.

23. Rodriguez-Wells, V., Plotch, S. J., and DeStefano, J. J. (2001) Primer-dependent synthesis by poliovirus RNA-dependent RNA polymerase (3Dpol). Nucleic Acids Res. 29, 2715-2724.

24. DeStefano, J. J., Critofaro, J. V., Derebail, S., Bohlayer, W. P., and Fitzgerald-Health, M. J. (2001) Physical mapping of HIV-RT to the 5' end of RNA primers. J. Biol. Chem. 276, 32515-32521.

25. Cristofaro J. V., Rausch, J. W., Le Grice, S. F. J., and DeStefano, J. J. (2002) Mutations in the RNase H active site of HIV-RT reveal a role for this site in stabilizing enzyme-primer-template binding. Biochemistry 41, 10968-10975.

26. Raja, A., and DeStefano, J. J. (2003) Interaction of HIV reverse transcriptase with structures mimicking recombination intermediates. J. Biol. Chem. 278, 10102-10111.

27. Liesch, G. R., and DeStefano, J. J. (2003) Analysis of mutations made during active synthesis or extension of mismatched substrates further define the mechanism of HIV-RT mutagenesis. Biochemistry 42, 5925-5936.

28. Derebail, S. S., Heath, M. J., and DeStefano, J. J. (2003) Evidence for the differential effects of nucleocapsid protein on strand transfer in various regions of the HIV genome. J. Biol. Chem. 278, 15702-15712.

29. Heath, M. J., Derebail, S. S., Gorelick, R. J., and DeStefano, J. J. (2003) Differing roles of the N-terminal and C-terminal zinc fingers in HIV-1 nucleocapsid protein enhanced nucleic acid annealing. J. Biol. Chem.278, 30755-30763.

30. Derebail S. S., and DeStefano J. J. (2004) Mechanistic analysis of pause site-dependent and -independent recombinogenic strand transfer from structurally diverse regions of the HIV genome. J. Biol. Chem. 279, 47446-47454.

31. Heath, M. J., and DeStefano, J. J. (2005) A complementary single stranded docking site is required for enhancement of strand exchange by human immunodeficiency virus nucleocapsid protein on substrates that model viral recombination. Biochemistry, 44, 3915-3925.

32. DeStefano, J. J. and Cristofaro, J. V. (2006) Selection of primer-template sequences that bind human immunodeficiency virus reverse transcriptase with high affinity. Nucleic Acids Res. 34:130-139.

33. DeStefano, J. J. and Odeyebo, T. (2006) Poliovirus protein 3AB displays nucleic acid chaperone and helix-destabilizing activities. J. Virol.80(4):1662-71.

34. Bohlayer W. P. and DeStefano J. J. (2006) Tighter binding of HIV reverse transcriptase to RNA-DNA vs. DNA-DNA results mostly from interactions in the polymerase domain and requires just a small stretch of RNA-DNA. Biochemistry 45(24):7628-38.

35. Narayanan, N, Gorelick, R. J., and DeStefano, J. J. (2006) Structure/function mapping of amino acids in the N-terminal zinc finger of the human immunodeficiency virus type 1 nucleocapsid protein: residues responsible for nucleic acid helix destabilizing activity.Biochemistry 45(41):12617-28.

36. Baird HA, Gao Y, Galetto R, Lalonde M, Anthony RM, Giacomoni V, Abreha M, Destefano JJ, Negroni M, Arts EJ. (2006) Influence of sequence identity and unique breakpoints on the frequency of intersubtype HIV-1 recombination. Retrovirology 3:91.

37. Anthony, R. M. and DeStefano J. J. (2007) In vitro synthesis of long DNA products in reactions with HIV-RT and nucleocapsid protein. J. Mol. Biol. 365(2):310-324.

38. DeStefano, J. J. and Nair, G. R. (2008) Novel aptamer inhibitors of human immunodeficiency virus reverse transcriptase. Oligonucleotides18(2):133-143.

39.     Jacob, D. T. and DeStefano J. J. (2008) A new role for HIV nucleocapsid protein in modulating the specificity of plus strand priming.Virology 378(2):385-396.

40.    DeStefano J. J. (2010) Effect of reaction conditions and 3AB on the mutation rate of poliovirus RNA-dependent RNA polymerase in a alpha-complementation assay.  Virus Res. 47(1):53-59.

41.   Olimpo, J. T. and DeStefano, J. J. (2010) Duplex structural differences and not 2’-hydroxyls explain the more stable binding of HIV-RT to RNA-DNA vs. DNA-DNA.  Nucleic Acids Res.38(10):4426-4435.

42.   Gangaramani, D. R., Eden, E. L., Shah, M., and DeStefano, J. J. (2010) The twenty-nine amino acid C-terminal cytoplasmic domain of poliovirus 3AB is critical for nucleic acid chaperone activity.  RNA Biology 7(6):8820-8829.

43.   Lai, Y. T., DeStefano J. J. (2011) A primer-free method that selects high-affinity single-stranded DNA aptamers using thermostable RNA ligase.  Anal. Biochem. 414(2):246-253.

44.   Fenstermacher, K. J., and DeStefano, J. J. (2011) Mechanism of HIV reverse transcriptase inhibition by zinc: formation of a highly stable enzyme-(primer-template) complex with profoundly diminished catalytic activity.  J. Biol. Chem. 286(47): 40433-40442.

45.   Lai, Y. T. and DeStefano, J. J. (2012) DNA aptamers to human immunodeficiency virus reverse transcriptase selected by a primer-free SELEX method: characterization and comparison to other aptamers.Nucleic Acids Therapeutics: 22(3):162-176.

46.   Nair, G.R., Dash, C., Le Grice, S.F. and DeStefano, J.J. (2012) Viral reverse transcriptases show selective high affinity binding to DNA-DNA primer-templates that resemble the polypurine tract. PloS One, 7, e41712.

47.   Lieberman O.J., DeStefano J.J. and Lee V.T. (2013) Detection of Cyclic Diguanylate G-Octaplex Assembly and Interaction with Proteins.PLoS ONE 8(1): e53689. doi:10.1371/journal.pone.0053689

48. Achuthan V., Keith B.J., Connolly B.A., DeStefano J.J. (2014) Human immunodeficiency virus reverse transcriptase displays dramatically higher fidelity under physiological magnesium conditions in vitro. J Virol. 88(15):8514-27. PMCID: PMC4135932.

49.  Achuthan V., DeStefano J.J. (2015) Alternative divalent cations (Zn2+, Co2+, and Mn2+) are not mutagenic at conditions optimal for HIV-1 reverse transcriptase activity. BMC Biochem. 16(1):12. PubMed PMID: 25934642.

50.  Miller MT, Tuske S, Das K, DeStefano JJ, Arnold E. (2015) Structure of HIV-1 reverse transcriptase bound to a novel 38-mer hairpin template-primer DNA aptamer. Protein Sci. 2015 Aug 21. doi: 10.1002/pro.2776. [Epub ahead of print] PubMed PMID: 26296781.

51.  Chauhan J, Chen SE, Fenstermacher KJ, Naser-Tavakolian A, Reingewertz T, Salmo R, Lee C, Williams E, Raje M, Sundberg E, DeStefano JJ, Freire E, Fletcher S. (2015) Synthetic, structural mimetics of the β-hairpin flap of HIV-1 protease inhibit enzyme function. Bioorg. Med. Chem. 23(21):7095-109. PMCID: PMC4661106.

52.  Alves Ferreira-Bravo I, Cozens C, Holliger P, DeStefano JJ. (2015) Selection of 2'-deoxy-2'-fluoroarabinonucleotide (FANA) aptamers that bind HIV-1 reverse transcriptase with picomolar affinity. Nucleic Acids Res. 16;43(20):9587-99. PubMed PMID: 26476448.

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