caa a22 4500 397494343 CHVBK 20180308164517.0 cr unu---uuuuu 161202e199503 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a124740 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a124740 Protein Factors Required for In Vitro Transcription of Sendai Virus Genome [Elektronische Daten] [Kiyohisa Mizumoto, Kenkoh Muroya, Toshimitsu Takagi, Toshiko Omata-Yamada, Hiroshi Shibuta, Kentaro Iwasaki] To elucidate the mechanism of transcription and replication of Sendai virus, we developed an efficient and faithful in vitro transcription system using purified virus particles. The in vitro RNA synthesis was almost entirely dependent on the addition of eukaryotic cell extracts, including those from various cultured mammalian cells, mammalian tissues, and even from plant cells. The RNA products were almost identical to authentic mRNA species synthesized in the infected cells, in their size distribution, the presence of 3′-poly(A) tail and the presence of methylated 5'-cap structure (m7GpppAm). Ribonuclease protection experiments after annealing the in vitro RNA with viral genomic RNA (vRNA) indicated that the virion-associated RNA-dependent RNA polymerase transcribes correct regions of the RNA genome in vitro. The active component(s) that is required for Sendai virus mRNA synthesis was partially purified from bovine brain and was separated into at least two complementary fractions, one of which could be replaced by highly purified cellular tubulin. When viral ribonucleoprotein complexes were used instead of virus particles in the in vitro transcription, only Sendai virus-infected cell extracts supported mRNA synthesis, and extracts from uninfected cells or cells infected with other viruses were found to be inert. These results suggest that, in addition to the general factors which are present ubiquitously in eukaryotic cells, a factor(s) specific to Sendai virus-infection is required for Sendai virus transcription. © 1995 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence cap structure nationallicence host factor nationallicence paramyxovirus nationallicence tubulin nationallicence viral ribonucleoprotein nationallicence Mizumoto Kiyohisa School of Pharmaceutical Sciences, Kitasato University Shirokane, Minato-ku, Tokyo 108 aut Muroya Kenkoh Institute of Medical Science, The University of Tokyo Shirokanedai, Minato-ku, Tokyo 108 aut Takagi Toshimitsu Institute of Medical Science, The University of Tokyo Shirokanedai, Minato-ku, Tokyo 108 aut Omata-Yamada Toshiko Tokyo Metropolitan Institute of Medical Science Honkomagome, Bunkyou-ku, Tokyo 113 aut Shibuta Hiroshi Institute of Medical Science, The University of Tokyo Shirokanedai, Minato-ku, Tokyo 108 aut Iwasaki Kentaro Tokyo Metropolitan Institute of Medical Science Honkomagome, Bunkyou-ku, Tokyo 113 aut The Journal of Biochemistry Oxford University Press 117/3(1995-03), 527-534 0021-924X 117:3<527 1995 117 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a124740 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a124740 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mizumoto Kiyohisa School of Pharmaceutical Sciences, Kitasato University Shirokane, Minato-ku, Tokyo 108 aut NATIONALLICENCE 700 1- Muroya Kenkoh Institute of Medical Science, The University of Tokyo Shirokanedai, Minato-ku, Tokyo 108 aut NATIONALLICENCE 700 1- Takagi Toshimitsu Institute of Medical Science, The University of Tokyo Shirokanedai, Minato-ku, Tokyo 108 aut NATIONALLICENCE 700 1- Omata-Yamada Toshiko Tokyo Metropolitan Institute of Medical Science Honkomagome, Bunkyou-ku, Tokyo 113 aut NATIONALLICENCE 700 1- Shibuta Hiroshi Institute of Medical Science, The University of Tokyo Shirokanedai, Minato-ku, Tokyo 108 aut NATIONALLICENCE 700 1- Iwasaki Kentaro Tokyo Metropolitan Institute of Medical Science Honkomagome, Bunkyou-ku, Tokyo 113 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 117/3(1995-03), 527-534 0021-924X 117:3<527 1995 117 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford