@article{Reich2014-11-11Initi-44422,
  year={2014},
  doi={10.1021/bi5006656},
  title={Initiation of RNA synthesis by the hepatitis C virus RNA-dependent RNA polymerase is affected by the structure of the RNA template},
  number={44},
  volume={53},
  issn={0006-2960},
  journal={Biochemistry},
  pages={7002--7012},
  author={Reich, Stefan and Kovermann, Michael and Lilie, Hauke and Knick, Paul and Geissler, René and Golbik, Ralph Peter and Balbach, Jochen and Behrens, Sven-Erik}
}

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    <dc:contributor>Lilie, Hauke</dc:contributor>
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    <dc:contributor>Golbik, Ralph Peter</dc:contributor>
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    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2019-01-07T12:01:49Z</dc:date>
    <dcterms:issued>2014-11-11</dcterms:issued>
    <dc:contributor>Kovermann, Michael</dc:contributor>
    <dc:contributor>Behrens, Sven-Erik</dc:contributor>
    <dc:contributor>Balbach, Jochen</dc:contributor>
    <dc:creator>Geissler, René</dc:creator>
    <dc:language>eng</dc:language>
    <dcterms:title>Initiation of RNA synthesis by the hepatitis C virus RNA-dependent RNA polymerase is affected by the structure of the RNA template</dcterms:title>
    <dc:creator>Knick, Paul</dc:creator>
    <dc:creator>Reich, Stefan</dc:creator>
    <dc:creator>Lilie, Hauke</dc:creator>
    <dc:contributor>Knick, Paul</dc:contributor>
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    <dc:contributor>Reich, Stefan</dc:contributor>
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    <dcterms:abstract xml:lang="eng">The hepatitis C virus (HCV) RNA-dependent RNA polymerase NS5B is a central enzyme of the intracellular replication of the viral (+)RNA genome. Here, we studied the individual steps of NS5B-catalyzed RNA synthesis by a combination of biophysical methods, including real-time 1D &lt;sup&gt;1&lt;/sup&gt;H NMR spectroscopy. NS5B was found to bind to a nonstructured and a structured RNA template in different modes. Following NTP binding and conversion to the catalysis-competent ternary complex, the polymerase revealed an improved affinity for the template. By monitoring the folding/unfolding of 3'(-)SL by &lt;sup&gt;1&lt;/sup&gt;H NMR, the base pair at the stem's edge was identified as the most stable component of the structure. &lt;sup&gt;1&lt;/sup&gt;H NMR real-time analysis of NS5B-catalyzed RNA synthesis on 3'(-)SL showed that a pronounced lag phase preceded the processive polymerization reaction. The presence of the double-stranded stem with the edge base pair acting as the main energy barrier impaired RNA synthesis catalyzed by NS5B. Our observations suggest a crucial role of RNA-modulating factors in the HCV replication process.</dcterms:abstract>
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    <dc:creator>Golbik, Ralph Peter</dc:creator>
    <dc:contributor>Geissler, René</dc:contributor>
    <dc:creator>Balbach, Jochen</dc:creator>
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    <dc:creator>Behrens, Sven-Erik</dc:creator>
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