title: Reconstitution of human rRNA gene transcription in mouse cells by a complete SL1 complex.
abstract: An important characteristic of the transcription of a ribosomal RNA gene (rDNA) mediated by DNA-dependent RNA polymerase (Pol) I is its stringent species specificity. SL1/TIF-IB is a key complex for species specificity, but its functional complex has not been reconstituted. Here, we established a novel and highly sensitive monitoring system for Pol I transcription to reconstitute the SL1 activity in which a transcript harboring a reporter gene synthesized by Pol I is amplified and converted into translatable mRNA by the influenza virus RNA-dependent RNA polymerase. Using this monitoring system, we reconstituted Pol I transcription from the human rDNA promoter in mouse cells by expressing four human TATA-binding protein (TBP)-associated factors (TAFIs) in the SL1 complex. The reconstituted SL1 also re-activated human rDNA transcription in mouse A9 cells carrying an inactive human chromosome 21 that contains the rDNA cluster. Chimeric SL1 complexes containing human and mouse TAFIs could be formed, but these complexes were inactive for human rDNA transcription. We conclude that four human TAFIs are necessary and sufficient to overcome the barrier of species specificity for human rDNA transcription in mouse cells.
authors.affiliation: Department of Infection Biology, Faculty of Medicine and Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Kensaku
authors.initials: K
authors.lastname: Murano
authors.affiliation: Department of Infection Biology, Faculty of Medicine and Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Okuwaki
authors.affiliation: Kitasato Institute for Life Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo 108-8641, Japan.
authors.forename: Fumitaka
authors.initials: F
authors.lastname: Momose
authors.affiliation: Department of Infection Biology, Faculty of Medicine and Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Michiko
authors.initials: M
authors.lastname: Kumakura
authors.affiliation: Department of Infection Biology, Faculty of Medicine and Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Shuhei
authors.initials: S
authors.lastname: Ueshima
authors.affiliation: Institute of Cancer Genetics and Pharmacogenomics, Division of Biosciences, School of Health Sciences and Social Care, Brunel University, Uxbridge, Middlesex UB8 3PH, UK.
authors.forename: Robert F
authors.initials: RF
authors.lastname: Newbold
authors.affiliation: Department of Infection Biology, Faculty of Medicine and Graduate School of Comprehensive Human Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki 305-8575, Japan knagata@md.tsukuba.ac.jp.
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Nagata
mesh_terms: D000818:Animals; D002460:Cell Line; D002904:Chromosomes, Human, 21-22 and Y / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D006801:Humans; D051379:Mice; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D009975:Orthomyxoviridae / Q000235:genetics*; D035463:Pol1 Transcription Initiation Complex Proteins / Q000378:metabolism*; D012318:RNA Polymerase I / Q000235:genetics / Q000378:metabolism*; D012335:RNA, Ribosomal / Q000235:genetics; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics / Q000378:metabolism*; D013045:Species Specificity; D035181:TATA-Box Binding Protein / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D020651:Ventral Thalamic Nuclei / Q000378:metabolism
chemical_list: D009687:Nuclear Proteins; D035463:Pol1 Transcription Initiation Complex Proteins; D012335:RNA, Ribosomal; D035181:TATA-Box Binding Protein; D014157:Transcription Factors; C063660:transcription initiation factor TIF-IB; C093517:transcriptional intermediary factor 1; D012324:RNA-Dependent RNA Polymerase; D012318:RNA Polymerase I
keywords: RNA polymerase I; Ribosomal RNA gene; SL1; Species specificity; TIF-IB; Transcription