Reconstitution of human rRNA gene transcription in mouse cells by a complete SL1 complex. 2014

Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
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.

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.

UI MeSH Term Description Entries
D009687 Nuclear Proteins Proteins found in the nucleus of a cell. Do not confuse with NUCLEOPROTEINS which are proteins conjugated with nucleic acids, that are not necessarily present in the nucleus. Nucleolar Protein,Nucleolar Proteins,Nuclear Protein,Protein, Nuclear,Protein, Nucleolar,Proteins, Nuclear,Proteins, Nucleolar
D009975 Orthomyxoviridae A family of RNA viruses causing INFLUENZA and other respiratory diseases. Orthomyxoviridae includes INFLUENZAVIRUS A; INFLUENZAVIRUS B; INFLUENZAVIRUS C; INFLUENZAVIRUS D; ISAVIRUS; and THOGOTOVIRUS. Influenza Viruses,Myxoviruses,Orthomyxoviruses,Influenza Virus,Myxovirus,Orthomyxovirus
D002460 Cell Line Established cell cultures that have the potential to propagate indefinitely. Cell Lines,Line, Cell,Lines, Cell
D002904 Chromosomes, Human, 21-22 and Y The short, acrocentric human chromosomes, called group G in the human chromosome classification. This group consists of chromosome pairs 21 and 22 and the Y chromosome. Chromosomes G,Group G Chromosomes,Chromosomes, Human, 21 22,Chromosomes, Human, 21-22,Chromosome, Group G,Chromosomes, Group G,Group G Chromosome
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012318 RNA Polymerase I A DNA-dependent RNA polymerase present in bacterial, plant, and animal cells. The enzyme functions in the nucleolar structure and transcribes DNA into RNA. It has different requirements for cations and salts than RNA polymerase II and III and is not inhibited by alpha-amanitin. DNA-Dependent RNA Polymerase I,RNA Polymerase A,DNA Dependent RNA Polymerase I,Polymerase A, RNA,Polymerase I, RNA
D012324 RNA-Dependent RNA Polymerase An enzyme that catalyses RNA-template-directed extension of the 3'- end of an RNA strand by one nucleotide at a time, and can initiate a chain de novo. (Enzyme Nomenclature, 1992, p293) Nucleoside-Triphosphate:RNA Nucleotidyltransferase (RNA-directed),RNA Replicase,RNA-Dependent RNA Replicase,RNA-Directed RNA Polymerase,RNA Dependent RNA Polymerase,RNA Dependent RNA Replicase,RNA Directed RNA Polymerase,RNA Polymerase, RNA-Dependent,RNA Polymerase, RNA-Directed,RNA Replicase, RNA-Dependent,Replicase, RNA,Replicase, RNA-Dependent RNA
D012335 RNA, Ribosomal The most abundant form of RNA. Together with proteins, it forms the ribosomes, playing a structural role and also a role in ribosomal binding of mRNA and tRNAs. Individual chains are conventionally designated by their sedimentation coefficients. In eukaryotes, four large chains exist, synthesized in the nucleolus and constituting about 50% of the ribosome. (Dorland, 28th ed) Ribosomal RNA,15S RNA,RNA, 15S
D013045 Species Specificity The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species. Species Specificities,Specificities, Species,Specificity, Species

Related Publications

Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
December 1994, Science (New York, N.Y.),
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
August 2005, Molecular and cellular biology,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
March 1995, European journal of biochemistry,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
January 2016, PloS one,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
April 2006, Cell cycle (Georgetown, Tex.),
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
March 1985, Molecular and cellular biology,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
December 1984, Philosophical transactions of the Royal Society of London. Series B, Biological sciences,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
November 1998, Journal of molecular biology,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
March 2001, The EMBO journal,
Kensaku Murano, and Mitsuru Okuwaki, and Fumitaka Momose, and Michiko Kumakura, and Shuhei Ueshima, and Robert F Newbold, and Kyosuke Nagata
January 2013, Standards in genomic sciences,
Copied contents to your clipboard!