title: Replication of partial double-stranded RNAs by Qβ replicase.
abstract: Qβ replicase, an RNA-dependent RNA polymerase of bacteriophage Qβ, uses single-stranded RNA as a template to synthesize the complementary strand. A single-stranded RNA template may contain rigid secondary structures, such as long stems, intermolecular double-stranded RNA regions. Presently, the effect of the size of such double-stranded regions on the replication of RNA by Qβ replicase is unknown. In this study, we prepared RNA templates hybridized with complementary RNA or DNA strands of various sizes and analyzed their replication by Qβ replicase. We found that Qβ replicase synthesizes the complementary strand as long as the template RNA is hybridized with no more than 200 nt fragments, although the replication amounts were decreased. This is important information to evaluate processivity of Qβ replicase.
authors.affiliation: Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Kenji
authors.initials: K
authors.lastname: Tomita
authors.affiliation: Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan; Exploratory Research for Advanced Technology, Japan Science and Technology Agency, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
authors.forename: Norikazu
authors.initials: N
authors.lastname: Ichihashi
authors.affiliation: Department of Bioinformatics Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan; Exploratory Research for Advanced Technology, Japan Science and Technology Agency, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan; Graduate School of Frontier Biosciences, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan. Electronic address: yomo@ist.osaka-u.ac.jp.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Yomo
mesh_terms: D017908:Allolevivirus / Q000201:enzymology / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D009693:Nucleic Acid Hybridization; D011777:Q beta Replicase / Q000235:genetics* / Q000378:metabolism; D012330:RNA, Double-Stranded / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D012330:RNA, Double-Stranded; D012367:RNA, Viral; D014764:Viral Proteins; D011777:Q beta Replicase
keywords: Double-stranded RNA; Qβ replicase; RNA duplex; RNA polymerization