In vitro Transcription of Escherichia coli Ribosomal RNA Genes

Nature ◽  
1972 ◽  
Vol 235 (5337) ◽  
pp. 329-333 ◽  
Author(s):  
WILLIAM A. HASELTINE
Keyword(s):  
Escherichia Coli ◽  
Ribosomal Rna ◽  
In Vitro Transcription ◽  
Ribosomal Rna Genes ◽  
Rna Genes

In vitro transcription of the ribosomal RNA genes of E. coli DNA

1977 ◽  
Vol 151 (3) ◽  
pp. 305-312 ◽  
Author(s):  
János Sümegi ◽  
Andor Udvardy ◽  
Pál Venetianer
Keyword(s):  
Ribosomal Rna ◽  
In Vitro Transcription ◽  
Ribosomal Rna Genes ◽  
E Coli ◽  
Rna Genes

Study of the function of Escherichia coli ribosomal RNA through site-directed mutagenesis

1990 ◽  
Vol 68 (1) ◽  
pp. 169-179 ◽  
Author(s):  
Daniel Leclerc ◽  
Léa Brakier-Gingras
Keyword(s):  
Escherichia Coli ◽  
Ribosomal Rna ◽  
Ribosomal Proteins ◽  
Large Body ◽  
Ribosomal Rna Genes ◽  
16S Ribosomal Rna ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Rna Genes

Various approaches have been developed to study how mutations in Escherichia coli ribosomal RNA affect the function of the ribosome. Most of them are in vivo approaches, where mutations are introduced in a specialized plasmid harboring the ribosomal RNA genes. The mutated plasmids are then expressed in an appropriate host, where they can confer resistance to antibiotics whose target is the ribosome. Conditions can be used where the host ribosomal RNA genes or the host ribosomes are selectively inactivated, and the effect of the mutations on ribosome assembly and function can be studied. Another approach, which has been developed mainly with 16S ribosomal RNA, can be used entirely in vitro. In this approach, a plasmid has been constructed which contains the 16S ribosomal RNA gene under control of a T7 promoter. Mutations can be introduced in the 16S ribosomal RNA sequence and the mutated 16S ribosomal RNAs are produced by in vitro transcription. It is then possible to investigate how the mutations affect the assembly of the 16S ribosomal RNA into 30S subunits and the activity of the reconstituted 30S subunits in cell-free protein synthesis assays. Although these approaches are recent, they have already provided a large body of interesting information, relating specific RNA sequences to interactions with ribosomal proteins, to ribosome function, and to its response to antibiotics.Key words: ribosomal RNA, ribosome, site-directed mutagenesis, antibiotic resistance.

scholarly journals Structural basis for the function of SuhB as a transcription factor in ribosomal RNA synthesis

2019 ◽  
Vol 47 (12) ◽  
pp. 6488-6503 ◽  
Author(s):  
Yong-Heng Huang ◽  
Nelly Said ◽  
Bernhard Loll ◽  
Markus C Wahl
Keyword(s):  
Escherichia Coli ◽  
Ribosomal Rna ◽  
Rna Synthesis ◽  
Rna Folding ◽  
In Vitro Transcription ◽  
Structural Basis ◽  
Macromolecular Crystallography ◽  
Rna Transcription ◽  
Signal Element

AbstractRibosomal RNA synthesis in Escherichia coli involves a transcription complex, in which RNA polymerase is modified by a signal element on the transcript, Nus factors A, B, E and G, ribosomal protein S4 and inositol mono-phosphatase SuhB. This complex is resistant to ρ-dependent termination and facilitates ribosomal RNA folding, maturation and subunit assembly. The functional contributions of SuhB and their structural bases are presently unclear. We show that SuhB directly binds the RNA signal element and the C-terminal AR2 domain of NusA, and we delineate the atomic basis of the latter interaction by macromolecular crystallography. SuhB recruitment to a ribosomal RNA transcription complex depends on the RNA signal element but not on the NusA AR2 domain. SuhB in turn is required for stable integration of the NusB/E dimer into the complex. In vitro transcription assays revealed that SuhB is crucial for delaying or suppressing ρ-dependent termination, that SuhB also can reduce intrinsic termination, and that SuhB-AR2 contacts contribute to these effects. Together, our results reveal functions of SuhB during ribosomal RNA synthesis and delineate some of the underlying molecular interactions.

scholarly journals Physical map of the seven ribosomal RNA genes of Escherichia coli

1979 ◽  
Vol 6 (5) ◽  
pp. 1817-1830 ◽  
Author(s):  
Imre Boros ◽  
Antal Kiss ◽  
Phá Venetianer
Keyword(s):  
Escherichia Coli ◽  
Ribosomal Rna ◽  
Physical Map ◽  
Ribosomal Rna Genes ◽  
Rna Genes

scholarly journals Transcription initiation of Xenopus 5S ribosomal RNA genes in vitro

1979 ◽  
Vol 7 (4) ◽  
pp. 947-958 ◽  
Author(s):  
Laurence J. Korn ◽  
Edward H. Birkenmeier ◽  
Donald D. Brown
Keyword(s):  
Ribosomal Rna ◽  
Transcription Initiation ◽  
Ribosomal Rna Genes ◽  
5S Ribosomal Rna ◽  
Rna Genes
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