| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
Animal: RNA Viruses |
Istituto di Ricerche di Biologia Molecolare ‘P. Angeletti’, via Pontina Km 30600, 00040-Pomezia (Roma), Italy1
Author for correspondence: Licia Tomei. Fax +39 06 91093225. e-mail Tomei{at}IRBM.it
The RNA-dependent RNA polymerase activity of hepatitis C virus is carried out by the NS5B protein. The full-length protein was previously purified as a non-fusion protein from insect cells infected with a recombinant baculovirus. The characterization is now described of a C-terminal hydrophobic domain deletion mutant of NS5B purified from E. coli. In addition to increased solubility, deletion of this sequence also positively affected the polymerase enzymatic activity. The efficiency of nucleotide polymerization of both the full-length and the C-terminal truncated enzymes were compared on homopolymeric template–primer couples as well as on RNA templates with heteropolymeric sequences. The largest difference in the polymerase activity was observed on the latter. On all the templates, the increased activity could be ascribed, at least in part, to enhanced template turnover of the deletion mutant with respect to the full-length enzyme. The elongation rates of the two enzyme forms were compared under single processive cycle conditions. Under these conditions, both the full-length and the deletion mutant were able to incorporate about 700 nt/min.
This article has been cited by other articles:
|
|
 |
|
  E. Ferrari, Z. He, R. E. Palermo, and H.-C. Huang Hepatitis C Virus NS5B Polymerase Exhibits Distinct Nucleotide Requirements for Initiation and Elongation J. Biol. Chem., December 5, 2008; 283(49): 33893 - 33901. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. A. Heck, A. M. I. Lam, N. Narayanan, and D. N. Frick Effects of Mutagenic and Chain-Terminating Nucleotide Analogs on Enzymes Isolated from Hepatitis C Virus Strains of Various Genotypes Antimicrob. Agents Chemother., June 1, 2008; 52(6): 1901 - 1911. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Kaushik-Basu, A. Bopda-Waffo, T. T. Talele, A. Basu, P. R. R. Costa, A. J. M. da Silva, S. G. Sarafianos, and F. Noel Identification and characterization of coumestans as novel HCV NS5B polymerase inhibitors Nucleic Acids Res., March 1, 2008; 36(5): 1482 - 1496. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 H. Lee, Y. Liu, E. Mejia, A. V. Paul, and E. Wimmer The C-Terminal Hydrophobic Domain of Hepatitis C Virus RNA Polymerase NS5B Can Be Replaced with a Heterologous Domain of Poliovirus Protein 3A J. Virol., November 15, 2006; 80(22): 11343 - 11354. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Kukolj, G. A. McGibbon, G. McKercher, M. Marquis, S. Lefebvre, L. Thauvette, J. Gauthier, S. Goulet, M.-A. Poupart, and P. L. Beaulieu Binding Site Characterization and Resistance to a Class of Non-nucleoside Inhibitors of the Hepatitis C Virus NS5B Polymerase J. Biol. Chem., November 25, 2005; 280(47): 39260 - 39267. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Di Marco, C. Volpari, L. Tomei, S. Altamura, S. Harper, F. Narjes, U. Koch, M. Rowley, R. De Francesco, G. Migliaccio, et al. Interdomain Communication in Hepatitis C Virus Polymerase Abolished by Small Molecule Inhibitors Bound to a Novel Allosteric Site J. Biol. Chem., August 19, 2005; 280(33): 29765 - 29770. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Y. M. Howe, J. Bloom, C. J. Baldick, C. A. Benetatos, H. Cheng, J. S. Christensen, S. K. Chunduru, G. A. Coburn, B. Feld, A. Gopalsamy, et al. Novel Nonnucleoside Inhibitor of Hepatitis C Virus RNA-Dependent RNA Polymerase Antimicrob. Agents Chemother., December 1, 2004; 48(12): 4813 - 4821. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Lee, H. Shin, E. Wimmer, and A. V. Paul cis-Acting RNA Signals in the NS5B C-Terminal Coding Sequence of the Hepatitis C Virus Genome J. Virol., October 15, 2004; 78(20): 10865 - 10877. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. A. van Dijk, E. V. Makeyev, and D. H. Bamford Initiation of viral RNA-dependent RNA polymerization J. Gen. Virol., May 1, 2004; 85(5): 1077 - 1093. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Liu, R. Chopra, S. Swanberg, S. Olland, J. O'Connell, and S. Herrmann Elongation of Synthetic RNA Templates by Hepatitis C Virus NS5B Polymerase J. Biol. Chem., March 12, 2004; 279(11): 10738 - 10746. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. McKercher, P. L. Beaulieu, D. Lamarre, S. LaPlante, S. Lefebvre, C. Pellerin, L. Thauvette, and G. Kukolj Specific inhibitors of HCV polymerase identified using an NS5B with lower affinity for template/primer substrate Nucleic Acids Res., January 22, 2004; 32(2): 422 - 431. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Tomei, S. Altamura, L. Bartholomew, M. Bisbocci, C. Bailey, M. Bosserman, A. Cellucci, E. Forte, I. Incitti, L. Orsatti, et al. Characterization of the Inhibition of Hepatitis C Virus RNA Replication by Nonnucleosides J. Virol., January 15, 2004; 78(2): 938 - 946. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. Tomei, S. Altamura, L. Bartholomew, A. Biroccio, A. Ceccacci, L. Pacini, F. Narjes, N. Gennari, M. Bisbocci, I. Incitti, et al. Mechanism of Action and Antiviral Activity of Benzimidazole-Based Allosteric Inhibitors of the Hepatitis C Virus RNA-Dependent RNA Polymerase J. Virol., December 15, 2003; 77(24): 13225 - 13231. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. J.-P. Leveque, R. B. Johnson, S. Parsons, J. Ren, C. Xie, F. Zhang, and Q. M. Wang Identification of a C-Terminal Regulatory Motif in Hepatitis C Virus RNA-Dependent RNA Polymerase: Structural and Biochemical Analysis J. Virol., August 15, 2003; 77(16): 9020 - 9028. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. W. Hardy, J. Marcotrigiano, K. J. Blight, J. E. Majors, and C. M. Rice Hepatitis C Virus RNA Synthesis in a Cell-Free System Isolated from Replicon-Containing Hepatoma Cells J. Virol., February 1, 2003; 77(3): 2029 - 2037. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Piccininni, A. Varaklioti, M. Nardelli, B. Dave, K. D. Raney, and J. E. G. McCarthy Modulation of the Hepatitis C Virus RNA-dependent RNA Polymerase Activity by the Non-Structural (NS) 3 Helicase and the NS4B Membrane Protein J. Biol. Chem., November 15, 2002; 277(47): 45670 - 45679. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. Labonte, V. Axelrod, A. Agarwal, A. Aulabaugh, A. Amin, and P. Mak Modulation of Hepatitis C Virus RNA-dependent RNA Polymerase Activity by Structure-based Site-directed Mutagenesis J. Biol. Chem., October 4, 2002; 277(41): 38838 - 38846. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Kim, H. Kim, S.-P. Cho, and M.-K. Min Template Requirements for De Novo RNA Synthesis by Hepatitis C Virus Nonstructural Protein 5B Polymerase on the Viral X RNA J. Virol., June 14, 2002; 76(14): 6944 - 6956. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Biroccio, J. Hamm, I. Incitti, R. De Francesco, and L. Tomei Selection of RNA Aptamers That Are Specific and High-Affinity Ligands of the Hepatitis C Virus RNA-Dependent RNA Polymerase J. Virol., March 19, 2002; 76(8): 3688 - 3696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sbardellati, E. Scarselli, E. Verschoor, A. De Tomassi, D. Lazzaro, and C. Traboni Generation of infectious and transmissible virions from a GB virus B full-length consensus clone in tamarins J. Gen. Virol., October 1, 2001; 82(10): 2437 - 2448. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Zhong, E. Ferrari, C. A. Lesburg, D. Maag, S. K. B. Ghosh, C. E. Cameron, J. Y. N. Lau, and Z. Hong Template/Primer Requirements and Single Nucleotide Incorporation by Hepatitis C Virus Nonstructural Protein 5B Polymerase J. Virol., October 1, 2000; 74(19): 9134 - 9143. [Abstract] [Full Text]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| INT J SYST EVOL MICROBIOL | MICROBIOLOGY | J GEN VIROL |
| J MED MICROBIOL | ALL SGM JOURNALS | |
