Monica Roth Professor UMDNJ-Robert Wood Johnson Medical School Department of Biochemistry Robert Wood Johnson Med School, Room 636 675 Hoes Lane Piscataway. NJ 08855 (732) 235-5048 FAX - 4783 roth@umdnj.edu

Retroviruses: integration. reverse transcriptase. envelope proteins. gene therapy. targeted entry. structural studies

Our research studies three stages of the retroviral life-cycle: entry. replication and integration. Retroviruses are RNA viruses capable of inducing various diseases including cancers and immunodeficiencies. The focus of our research is to study three stages in the infectious retroviral life cycle. These include the entry. replication and integration of the virus.

Entry: Retroviral entry is defined by the interaction of the host receptor protein with the viral envelope proteins. Retroviruses are frequently used as a means of gene delivery for gene therapy. We have developed a new approach to retarget the retroviral entry to novel receptors using a mammalian Env random display library. This system is now being applied to biologically relevant systems including cancer cells. The ability to specifically target viral entry into selective cells has widespread applications.

Additional studies aim at understanding on the molecular level the mechanism by which Murine Leukemia Viruses choose and enter their host cells. Viral gene products are known to be required for the entry of the virus into cells. In our laboratory these proteins. the Envelope gene products. are studied genetically and biochemically.

Replication and Integration: The retroviral pol gene encodes the reverse transcriptase (RT) and integrase (IN) proteins. Our research involves the expression. purification and characterization of the reverse transcriptase and integrase proteins. The reverse transcriptase is a multifunctional protein. which replicates the viral RNA into double-stranded DNA form. This DNA product is subsequently integrated into the host DNA. a reaction requiring the viral integrase. A main focus of the studies on viral integration involve the X-ray crystallographic analysis of the MuLV integrase protein and subdomains.

Selected Publications

Mazari PM, Linder-Basso D, Sarangi A, Chang Y, Roth MJ. (2009) Single-round selection yields a unique retroviral envelope utilizing GPR172A as its host receptor. Proc Natl Acad Sci U S A. 106(14):5848-53.

Vera J, Valenzuela B, Roth MJ, León O. (2008) Characterization of the long-terminal repeat single-strand tail-binding site of Moloney-MuLV integrase by crosslinking. Biol Res. 41(1):69-80.

Coté ML, Roth MJ.(2008) Murine leukemia virus reverse transcriptase: structural comparison with HIV-1 reverse transcriptase. Virus Res. 134(1-2):186-202.

Schneider WM, Zheng H, Coté ML, Roth MJ. (2008) The MuLV 4070A G541R Env mutation decreases the stability and alters the conformation of the TM ectodomain. Virology. 371(1):165-74.

Sarangi A, Bupp K, Roth MJ. (2007) Identification of a retroviral receptor used by an envelope protein derived by peptide library screening. Proc Natl Acad Sci U S A. 104(26):11032-7.

Montano SP, Cote ML, Roth MJ, Georgiadis MM. (2006) Crystal structures of oligonucleotides including the integrase processing site of the Moloney murine leukemia virus. Nucleic Acids Res. 34(19):5353-60.

Puglia J, Wang T, Smith-Snyder C, Cote M, Scher M, Pelletier JN, John S, Jonsson CB, Roth MJ. (2006) Revealing domain structure through linker-scanning analysis of the murine leukemia virus (MuLV) RNase H and MuLV and human immunodeficiency virus type 1 integrase proteins. J Virol. 80(19):9497-510.

Bupp K. Sarangi A. Roth MJ. (2006) Selection of feline leukemia virus envelope proteins from a library by functional association with a murine leukemia virus envelope. Virology. 351(2):340-8.

Bupp K. Roth MJ. (2005) Alteration and analyses of viral entry with library-derived peptides. Adv Virus Res. 65:147-72.

Bupp. K. Sarangi. A. and Roth. MJ (2005) Probing sequence variation in the receptor-targeting domain of feline leukemia virus envelope proteins using peptide display libraries. J. Virol. 79:1463-1469.

Sunigo. R. Sengupta. S. Snyder. C. Leon. O and Roth. MJ(2004) Expression of the C-terminus of HIV-1 reverse transcriptase P66 and P51 subunits as a single polypeptide with RNase H activity. Protein Engineering. Design and Selection 17: 581-587. (Cover)

Guaitiao. JP. Zúñiga. RA. Roth. MJ and Leon. O (2004) Lysine directed crosslinking of viral RNA/DNA hybrid substrate to the isolated RNase H domain of HIV-1 reverse transcriptase. Biochemistry 43: 1302-8.

Bupp K and Roth. MJ (2003) Targeting a retroviral vector in the absence of a known cell-targeting ligand. Human Gene Therapy 14(16) 1557-64.

Villanueva. RA. Campbell. S. and Roth. MJ (2003) Molecular analysis of a recombinant M-MuLV-RaLV retrovirus. Virology 314:195-208.

Villanueva. RA. Jonsson. CB. Jones. J. Georgiadis. MM and Roth. MJ (2003) Differential multimerization of Moloney murine leukemia virus integrase purified under non-denaturing conditions. Virology 316:146-160.

O'Reilly. L and Roth. MJ (2003) G541R within the 4070A TM protein regulates fusion in Murine Leukemia Viruses. J. Virol 77:12011-12021.

Lu. C. -W. . O'Reilly. L. and Roth. M. J. (2003) G100R mutation within 4070A MuLV Env increases virus receptor binding. kinetics of entry. and viral transduction efficiency. J. Virol:77:739-743.

Lu. C.-W. and Roth. M. J. (2003) Functional interaction between the N- and C-terminaldomains of murine leukemia virus surface Envelope protein. Virology:310:130 - 140.

O'Reilly. L. and Roth. M. J. (2003) Identification of conformational and cold-sensitive mutations in the MuLV Envelope protein. Virology:312:337-49.

Yang. F. . Belitsky. J. M. . Villanueva. R. A.. Dervan. P. B. and Roth. M. J. (2003) Inhibition of Moloney murine leukemia virus integration using polyamides targeting the long-terminal repeat (LTR) sequences. Biochemistry 42(20):6249-58.

Lu. C. -W. and Roth. M. J. (2003) The role of Q252R in activating membrane fusion in the murine leukemia virus surface Envelope protein. J. Virol. 77: 10841-10849.

Bupp. K. and Roth. M. J. (2002) Altering retroviral tropism using a random display Envelope library. Molecular Therapy 5:329-335.

Seamon. J. A.. Jones. K. S.. Miller. C. and Roth. M. J. (2002) Inserting a nuclear targeting signal onto a replication-competent M-MuLV affects viral export and is not sufficient for cell cycle independent infection. J. Virol. 76: 8475-8484.

Yang. F. and Roth. M. J. (2001) Assembly and catalysis of concerted two-end integration events by Moloney murine leukemia virus. J. Virol 75:9561-9570.

Yang. F. . Seamon. J. A. and Roth. M. J. (2001) Mutational analysis of the N-terminus of Moloney murine leukemia virus integrase. Virology 291:32-45.

Gu. J.. Villanueva. R. A.. Synder. C. S.. Roth. M. J. and Georgiadis. M. M. (2001) Substitution of Asp 114 or Arg 116 in the fingers domain of Moloney murine leukemia virus reverse transcriptase affects interactions with the primer-template resulting in decreased processivity. J. Mol. Biol. 305:341-359.

Lu. C.-W. and Roth. M. J. (2001) Functional characterization of the N-terminus of murine leukemia virus Envelope proteins. J. Virol. 72:4357-4366.

Snyder. C.S. and Roth. M. J. (2001) Retroviral RNase H. Methods in Enzymology. (Academic Press. San Diego. CA) Allen W. Nicholson. editor. Vol.341 Part A. pp. 440-452.