University of Medicine and Dentistry of New Jersey

Tahirov,T.; Temiakov,D.; Anikin,M.; Patlan,V.; McAllister,W.T.; Vasylyev,D.G.; Yokoyama,S. Structure of a T7 RNA polmerase elongation complex at 2.9A resolution. Nature. Vol. 420: 43-50 (2002 ) .

Kukarin, A., Rong, M.R., McAllister, W.T. Exposure of T7 TNA polymerase to the double stranded binding region of the promoter activates the enzyme to transcribe a single stranded template. J. Biol. Chem. Vol. 278: 2419-2424 (2003 ) .

Temiakov, D., Patlan, V., Anikin, M., McAllister, W.T., Yokoyama S., Vassylyev, D.G. Structural basis for substrate selection by T7 RNA polymerase. Cell. Vol. 116: 381-391 (2004 ) .

Jiang, M., Ma, N., Vassylyev, D.G., McAllister, W.T. RNA displacement and resolution of the transcription bubble during transcription by T7 RNA polymerase. Mol. Cell. Vol. 15: 777-788 (2004 ) .

Pomerantz, R.T., Ramjit, R., Gueroui, Z., Place, C., Anikin, M., Leuba, S., Zlatanova, J., McAllister, W.T. A Tightly Regulated Molecular Motor Based upon T7 RNA Polymerase. Nanno Letters. Vol. 5: 1698-1703 (2005 ) .

Ma K, Temiakov D, Anikin M, McAllister WT Probing conformational changes in T7 RNA polymerase during initiation and termination by using engineered disulfide linkages. Proc Natl Acad Sci U S A. Vol. 2005 Dec 6;102(49): 17612-7 Epub 2005 Nov 21 (2005 ) .

Zlatanova J, McAllister WT, Borukhov S, Leuba SH. Single-molecule approaches reveal the idiosyncrasies of RNA polymerases. Structure.. Vol. 2006 Jun;14(6): 953-66 (2006 ) .

Pomerantz, R.T., Temiakov, D., Anikin, M., Vassylev, D.G., McAllister, W.T. A novel mechanism of nucleotide misincorporation during transcription due to template strand misalignment. Mol. Cell. Vol. 24: 245-255 (2006 ) .

Kashkina, E., Anikin, M., Bruckner, F., Pomerantz, R.T., McAllister, W.T., Cramer, P., Temiakov, D. Template misalignment in multisubunit RNA polymerases and transciption fidelity. Mol. Cell. Vol. 24: 256-257 (2006 ) .

Kashkina, E., Anikin, M. Brueckner, F., Lehmann, E., Kochetkov, S.N., McAllister, W.T., Cramer, P., Temiakov, T. Multi-subunit RNA polymerases mel only a single DNA base pair downstream of the active site. J. Biol. Chem.. Vol. 30: 2158-2182 (2007 ) .

Bandwar, R.P., Ma, N., Emanuel, S.A., Anikin, M., Vassylyev, D.G., Patel, S.S., McAllister, W.T. The transition to an elongation complex by T7 RNA polymerase is a multistep process. J. Biol. Chem.. Vol. 31: 22879-22886 (2007 ) .

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Grants and Contracts

Title: "RNA polymerase structure and function"
Sponsor: NIH
Effective Date(s): 2004 - 2008
Role: Principal Investigator

Title: "Structure-function relationships of bacteriophage T7 RNA polymerase"
Sponsor: Howard Hughes Medical Institute
Effective Date(s): 1995 - 2001
Role: Co-Investigator

Title: "RNA polymerases with altered specificities"
Sponsor: bioMerieux, S.A.
Effective Date(s): 1993 - 1998
Role: Principal Investigator

Title: "Development of phage RNA polymerase-based expression system"
Sponsor: Life Technologies, Inc.
Effective Date(s): 1992 - 1994
Role: Principal Investigator

Title: "Regulation of viral gene expression"
Sponsor: NIH
Effective Date(s): 1974 - 1988
Role: Principal Investigator

Title: "Studies of recombination in papillomavirus-transformed cells"
Sponsor: New Jersey Commission on Cancer Research
Effective Date(s): 1985 - 1987
Role: Principal Investigator

Title: "Support for an international workshop on gene organization and expression in bacteriophages"
Sponsor: NSF
Effective Date(s): 1988 - 1989
Role: Principal Investigator

Title: "Support of an International Workshop on Macromolecular Interactions in Bacteriophages"
Sponsor: NSF
Effective Date(s): 1996 - 1997
Role: Principal Investigator

Title: "Cloning and expression of the bacteriophage SP6 RNA polymerase gene"
Sponsor: Pharmacia P-L Biochemicals, Inc.
Effective Date(s): 1987 - 1990
Role: Principal Investigator

Title: "Development of phage RNA polymerase-based expression systems"
Sponsor: Life Technologies, Inc.
Effective Date(s): 1991 - 1994
Role: Principal Investigator

Title: "Development of a plant expression system based upon phage T3 RNA polymerase"
Sponsor: Biotechnology Research and Development Corporation
Effective Date(s): 1992 - 1993
Role: Principal Investigator

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Research Areas

Research Interests:: Work in our laboratory is directed at understanding fundamental aspects of the transcription process, and in particular, how the enzyme that carries out this process works. As a model system we have chosen the relatively simple RNA polymerase (RNAP) that is encoded by bacteriophage T7. Although this enzyme consists of a single subunit, it carries out all of the steps in the transcription process in a similar manner as the multisubunit RNAPs found in prokaryotic and eukaryotic cells. Importantly, the phage enzyme is structurally related to other nucleotide polymerases of the pol I family, such as DNA polymerase I and reverse transcriptase. It therefore affords an opportunity to explore and compare general features of nucleotide polymerization and fidelity among members of this family, as well as with multisubunit enzymes. In our work we use biochemical and genetic methods to characterize the importance of various regions of the RNAP to RNA synthesis, and interpret the results with regard to the organization of the enzyme as determined by crystallographic analysis and other methods of structural determination. A number of structures of T7 RNAP have been solved, including: free RNAP, RNAP bound to the promoter, an early initiation complex, and, most recently, the structure of an elongation complex. These structures, together with structures of bacterial and yeast RNAPs, have provided a wealth of information concerning common features of the transcription machinery, and important insights into the transcription process. Nevertheless, important gaps remain in our knowledge of the various stages of transcription, most importantly with regard to the transition that leads from an unstable initiation complex (IC) to a stable elongation complex (EC), and with regard to the process of termination.

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