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dc.creatorLee, Heng-Chien
dc.creatorAalto, Antti P.en
dc.creatorYang, Qiuyingen
dc.creatorChang, Shwu-Shinen
dc.creatorHuang, Guocunen
dc.creatorFisher, Danielen
dc.creatorCha, Joonseoken
dc.creatorPoranen, Minna M.en
dc.creatorBamford, Dennis H.en
dc.creatorLiu, Yien
dc.date.accessioned2013-06-28T15:18:51Zen
dc.date.available2013-06-28T15:18:51Zen
dc.date.issued2010-10-05en
dc.identifier.citationLee H-C, Aalto AP, Yang Q, Chang S-S, Huang G, et al. (2010) The DNA/RNA-Dependent RNA Polymerase QDE-1 Generates Aberrant RNA and dsRNA for RNAi in a Process Requiring Replication Protein A and a DNA Helicase. PLoS Biol 8(10): e1000496. doi:10.1371/journal.pbio.1000496en
dc.identifier.urihttp://hdl.handle.net/2152/20521en
dc.descriptionHeng-Chi Lee is with the University of Texas Southwestern Medical Center, Antti P. Aalto is with University of Helsinki, Qiuying Yang is with the University of Texas Southwestern Medical Center, Shwu-Shin Chang is with the University of Texas Southwestern Medical Center, Guocun Huang is with the University of Texas Southwestern Medical Center, Daniel Fisher is with the University of Texas Southwestern Medical Center and UT Austin, Joonseok Cha is with the University of Texas Southwestern Medical Center, Minna M. Poranen is with University of Helsinki, Dennis H. Bamford is with University of Helsinki, Yi Liu is with the University of Texas Southwestern Medical Center.en
dc.description.abstractThe production of aberrant RNA (aRNA) is the initial step in several RNAi pathways. How aRNA is produced and specifically recognized by RNA-dependent RNA polymerases (RdRPs) to generate double-stranded RNA (dsRNA) is not clear. We previously showed that in the filamentous fungus Neurospora, the RdRP QDE-1 is required for rDNA-specific aRNA production, suggesting that QDE-1 may be important in aRNA synthesis. Here we show that a recombinant QDE-1 is both an RdRP and a DNA-dependent RNA polymerase (DdRP). Its DdRP activity is much more robust than the RdRP activity and occurs on ssDNA but not dsDNA templates. We further show that Replication Protein A (RPA), a single-stranded DNA-binding complex that interacts with QDE-1, is essential for aRNA production and gene silencing. In vitro reconstitution assays demonstrate that QDE-1 can produce dsRNA from ssDNA, a process that is strongly promoted by RPA. Furthermore, the interaction between QDE-1 and RPA requires the RecQ DNA helicase QDE-3, a homolog of the human Werner/Bloom Syndrome proteins. Together, these results suggest a novel small RNA biogenesis pathway in Neurospora and a new mechanism for the production of aRNA and dsRNA in RNAi pathways.en
dc.description.sponsorshipThis study was supported by grants from National Institutes of Health and Welch Foundation (I-1560) to YL, and the Academy of Finland Finnish Centre of Excellence Programme 2006-2011 (1129684) to DB. AA is supported by the Helsinki Graduate School in Biotechnology and Molecular Biology. YL is the Louise W. Kahn endowed scholar in Biomedical Research at University of Texas Southwestern Medical Center. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.en
dc.language.isoengen
dc.publisherPublic Library of Scienceen
dc.rightsAttribution 3.0 United Statesen
dc.rightsCC-BYen
dc.rights.urihttp://creativecommons.org/licenses/by/3.0/us/en
dc.subjectDNA damageen
dc.subjectDNA-RNA hybridizationen
dc.subjectGenetic locien
dc.subjectNeurosporaen
dc.subjectRNA denaturationen
dc.subjectRNA polymeraseen
dc.subjectRNA synthesisen
dc.subjectSmall interfering RNAen
dc.titleThe DNA/RNA-Dependent RNA Polymerase QDE-1 Generates Aberrant RNA and dsRNA for RNAi in a Process Requiring Replication Protein A and a DNA Helicaseen
dc.typeArticleen
dc.description.departmentPsychologyen
dc.identifier.doi10.1371/journal.pbio.1000496en


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Attribution 3.0 United States
Except where otherwise noted, this item's license is described as Attribution 3.0 United States