PMID: 8948635Nov 15, 1996Paper

Identification of two separate domains in the influenza virus PB1 protein involved in the interaction with the PB2 and PA subunits: a model for the viral RNA polymerase structure

Nucleic Acids Research
S GonzálezJ Ortín

Abstract

The domains of the PB1 subunit of the influenza virus polymerase involved in the interaction with the PB2 and PA subunits have been defined by mutational analysis of PB1 protein. The experimental approach included in vivo competition of the PB1 activity, two-hybrid interaction assays and in vitro binding to PB1-specific matrices. Mutants of the PB1 gene including N-terminal, C-terminal and internal deletions and single amino acid insertions were constructed. They were unable to support polymerase activity in a reconstituted transcription-replication system and were tested for their competition activity when expressed in excess over wild-type PB1 protein. The pattern of competition obtained suggested that the N-terminal 78 amino acids and the sequences between positions 506 and 659 in the PB1 protein are involved in the interaction with the other components of the polymerase. We identified the N-terminal region of PB1 protein as responsible for the interaction with the PA subunit by two-hybrid assays in mammalian cells. N- and C-terminal fragments of the PB1 protein were expressed as His-tagged proteins and purified on Ni2+-NTA resin. Such PB1-specific matrices were used in binding assays in vitro with metabolically labelled PB2...Continue Reading

References

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Citations

Oct 12, 2012·Archives of Virology·Yanfeng YaoZe Chen
Oct 20, 2012·Bulletin of Mathematical Biology·Alex MadrahimovJim Rogers
Feb 22, 2008·Proceedings of the National Academy of Sciences of the United States of America·Claudia PappasTerrence M Tumpey
Apr 5, 2012·Proceedings of the National Academy of Sciences of the United States of America·Giulia MuratoreArianna Loregian
Aug 12, 2010·The Journal of Biological Chemistry·Atsushi IwaiTadaaki Miyazaki
Jun 12, 2010·The Journal of Biological Chemistry·Stéphane BoivinDarren J Hart
May 23, 2007·Nucleic Acids Research·Eva TorreiraOscar Llorca
May 15, 2002·Microbiology and Immunology·Yasushi OhtsuTetsuya Toyoda
May 12, 2007·Journal of Virology·Alexander GhanemMartin Schwemmle
Jun 15, 2012·Journal of Virology·Dorothée MoisyJean-Luc Jestin
Apr 24, 2009·Annual Review of Microbiology·Terrence M Tumpey, Jessica A Belser
May 6, 2010·Future Microbiology·Nikolai Kaverin
Dec 24, 2003·Proceedings of the National Academy of Sciences of the United States of America·Estela AreaJuan Ortín
Apr 5, 2014·Cellular and Molecular Life Sciences : CMLS·Arianna LoregianGiorgio Palù
Jun 12, 2009·Protein Science : a Publication of the Protein Society·Mathias DregerTao Deng

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