HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 276, Issue 33, 31179-31185, August 17, 2001

This Article Full Text Full Text (PDF) All Versions of this Article: 276/33/31179    most recent M102856200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Honda, A. Articles by Ishihama, A. Search for Related Content PubMed PubMed Citation Articles by Honda, A. Articles by Ishihama, A. Social Bookmarking                      What's this?

Differential Roles of Viral RNA and cRNA in Functional Modulation of the Influenza Virus RNA Polymerase^*

Ayae Honda^§¶, Atsushi Endo, Kiyohisa Mizumoto^**, and Akira Ishihama

From the  Department of Molecular Genetics, National Institute of Genetics, Mishima, Shizuoka 411-8540, Japan, the ^§ Japan Science and Technology Corp., Kawaguchi, Saitama 332-0012, Japan, the  Daiichi Pharmaceutical Co., Ltd., Exploratory Research Laboratories, Edogawa, Tokyo 134-0081, Japan, and the ^** Faculty of Pharmaceutical Science, Kitasato University, Minato-ku, Tokyo 108-8641, Japan

The RNA-dependent RNA polymerase of influenza virus is composed of three viral P proteins (PB1, PB2, and PA) and involved in both transcription and replication of the RNA genome. For the molecular anatomy of this multifunctional enzyme, we have established a simultaneous expression of three P proteins in cultured insect cells using recombinant baculoviruses. For purification of P protein complexes, the PA protein was expressed as a fusion with a histidine tag added at its N terminus. By using affinity chromatography, a complex consisting of the three P proteins was isolated from nuclear extracts of virus-infected cells. The affinity-purified 3P complex showed the activities of capped RNA binding, capped RNA cleavage, viral model RNA binding, model RNA-directed RNA synthesis, and polyadenylation of newly synthesized RNA. We conclude that a functional form of the viral RNA polymerase with the catalytic specificity of transcriptase is formed in recombinant baculovirus-infected insect cells. Using the viral RNA-free 3P complex, we found that the capped RNA cleavage takes place in the presence of vRNA but not of cRNA, indicating that the vRNA functions as a regulatory factor for the specificity control of viral RNA polymerase as well as a template for transcription. The structural elements of RNA directing the expression of RNA polymerase functions were analyzed using variant forms of the model RNA templates.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				A. Honda, T. Okamoto, and A. Ishihama Host factor Ebp1: Selective inhibitor of influenza virus transcriptase. Genes Cells, February 1, 2007; 12(2): 133 - 142. [Abstract] [Full Text] [PDF]

				M. Crow, T. Deng, M. Addley, and G. G. Brownlee Mutational Analysis of the Influenza Virus cRNA Promoter and Identification of Nucleotides Critical for Replication J. Virol., June 15, 2004; 78(12): 6263 - 6270. [Abstract] [Full Text] [PDF]

				A. Honda, K. Mizumoto, and A. Ishihama Minimum molecular architectures for transcription and replication of the influenza virus PNAS, October 1, 2002; 99(20): 13166 - 13171. [Abstract] [Full Text] [PDF]

				G. G. Brownlee and J. L. Sharps The RNA Polymerase of Influenza A Virus Is Stabilized by Interaction with Its Viral RNA Promoter J. Virol., June 14, 2002; 76(14): 7103 - 7113. [Abstract] [Full Text] [PDF]

				M. T. M. Lee, K. Bishop, L. Medcalf, D. Elton, P. Digard, and L. Tiley Definition of the minimal viral components required for the initiation of unprimed RNA synthesis by influenza virus RNA polymerase Nucleic Acids Res., January 15, 2002; 30(2): 429 - 438. [Abstract] [Full Text] [PDF]