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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
Insights into the pre-initiation events of bacteriophage φ6 RNA-dependent RNA polymerase: Towards the assembly of a productive binary complex
Nucleic Acids Research, Volume 37, No. 4, Year 2009
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Description
The RNA-dependent RNA polymerase (RdRP) of double-stranded RNA (dsRNA) viruses performs both RNA replication and transcription. In order to initiate RNA polymerization, viral RdRPs must be able to interact with the incoming 3′ terminus of the template and position it, so that a productive binary complex is formed. Structural studies have revealed that RdRPs of dsRNA viruses that lack helicases have electrostatically charged areas on the polymerase surface, which might facilitate such interactions. In this study, structure-based mutagenesis, enzymatic assays and molecular mapping of bacteriophage φ6 RdRP and its RNA were used to elucidate the roles of the negatively charged plough area on the polymerase surface, of the rim of the template tunnel and of the template specificity pocket that is key in the formation of the productive RNA-polymerase binary complex. The positively charged rim of the template tunnel has a significant role in the engagement of highly structured ssRNA molecules, whereas specific interactions further down in the template tunnel promote ssRNA entry to the catalytic site. Hence, we show that by aiding the formation of a stable binary complex with optimized RNA templates, the overall polymerization activity of the φ6 RdRP can be greatly enhanced. © 2009 The Author(s).
Available Materials
https://efashare.b-cdn.net/share/pmc/articles/PMC2651803/bin/gkn1035_index.html
https://efashare.b-cdn.net/share/pmc/articles/PMC2651803/bin/gkn1035_1.pdf
https://efashare.b-cdn.net/share/pmc/articles/PMC2651803/bin/gkn1035_2.pdf
https://efashare.b-cdn.net/share/pmc/articles/PMC2651803/bin/gkn1035_3.pdf
Authors & Co-Authors
Sarin, L. Peter
Finland, Helsinki
Helsingin Yliopisto
Poranen, Minna M.
Finland, Helsinki
Helsingin Yliopisto
Aalto, Antti P.
Finland, Helsinki
Helsingin Yliopisto
Van Dijk, A. A.
Finland, Helsinki
Helsingin Yliopisto
Stuart, David Ian
United Kingdom, Oxford
University of Oxford
Grimes, Jonathan M.
United Kingdom, Oxford
University of Oxford
Bamford, Dennis Henry
Finland, Helsinki
Helsingin Yliopisto
Statistics
Citations: 26
Authors: 7
Affiliations: 2
Identifiers
Doi:
10.1093/nar/gkn1035
ISSN:
13624962
Research Areas
Genetics And Genomics