Characterisation of heparin-resistant complex formation and RNA synthesis by wheat germ RNA polymerases I, II and III, in vitro on cauliflower mosaic virus DNA

1981 ◽  
Vol 98 (1) ◽  
pp. 36-42 ◽  
Author(s):  
R. Cooke ◽  
R. Durand ◽  
M. Teissere ◽  
P. Penon ◽  
J. Ricard
Keyword(s):  
Complex Formation ◽  
Mosaic Virus ◽  
Wheat Germ ◽  
Rna Synthesis ◽  
Rna Polymerases ◽  
Cauliflower Mosaic Virus

scholarly journals Role of the 3′ tRNA-Like Structure in Tobacco Mosaic Virus Minus-Strand RNA Synthesis by the Viral RNA-Dependent RNA Polymerase In Vitro

2000 ◽  
Vol 74 (24) ◽  
pp. 11671-11680 ◽  
Author(s):  
T. A. M. Osman ◽  
C. L. Hemenway ◽  
K. W. Buck
Keyword(s):  
Rna Polymerase ◽  
Tobacco Mosaic Virus ◽  
Mosaic Virus ◽  
Untranslated Region ◽  
Rna Synthesis ◽  
Central Core ◽  
Minus Strand ◽  
Stem Loop ◽  
Polymerase Binding

ABSTRACT A template-dependent RNA polymerase has been used to determine the sequence elements in the 3′ untranslated region of tobacco mosaic virus RNA that are required for promotion of minus-strand RNA synthesis and binding to the RNA polymerase in vitro. Regions which were important for minus-strand synthesis were domain D1, which is equivalent to a tRNA acceptor arm; domain D2, which is similar to a tRNA anticodon arm; an upstream domain, D3; and a central core, C, which connects domains D1, D2, and D3 and determines their relative orientations. Mutational analysis of the 3′-terminal 4 nucleotides of domain D1 indicated the importance of the 3′-terminal CA sequence for minus-strand synthesis, with the sequence CCCA or GGCA giving the highest transcriptional efficiency. Several double-helical regions, but not their sequences, which are essential for forming pseudoknot and/or stem-loop structures in domains D1, D2, and D3 and the central core, C, were shown to be required for high template efficiency. Also important were a bulge sequence in the D2 stem-loop and, to a lesser extent, a loop sequence in a hairpin structure in domain D1. The sequence of the 3′ untranslated region upstream of domain D3 was not required for minus-strand synthesis. Template-RNA polymerase binding competition experiments showed that the highest-affinity RNA polymerase binding element region lay within a region comprising domain D2 and the central core, C, but domains D1 and D3 also bound to the RNA polymerase with lower affinity.

Brome mosaic virus coat protein inhibits viral RNA synthesis in vitro

Virology ◽  
1987 ◽  
Vol 158 (1) ◽  
pp. 15-19 ◽  
Author(s):  
Mamoru Horikoshi ◽  
Masaharu Nakayama ◽  
Naoto Yamaoka ◽  
Iwao Furusawa ◽  
Jiko Shishiyama
Keyword(s):  
Coat Protein ◽  
Mosaic Virus ◽  
Rna Synthesis ◽  
Brome Mosaic Virus ◽  
Viral Rna ◽  
Virus Coat Protein ◽  
Virus Coat

Role of the Pepino mosaic virus 3′-untranslated region elements in negative-strand RNA synthesis in vitro

2014 ◽  
Vol 190 ◽  
pp. 110-117 ◽  
Author(s):  
Toba A.M. Osman ◽  
René C.L. Olsthoorn ◽  
Ioannis C. Livieratos
Keyword(s):  
Mosaic Virus ◽  
Untranslated Region ◽  
Rna Synthesis ◽  
Pepino Mosaic Virus ◽  
Negative Strand

Mutational studies of archaeal RNA polymerase and analysis of hybrid RNA polymerases

2009 ◽  
Vol 37 (1) ◽  
pp. 18-22 ◽  
Author(s):  
Michael Thomm ◽  
Christoph Reich ◽  
Sebastian Grünberg ◽  
Souad Naji
Keyword(s):  
Complex Formation ◽  
Structural Similarity ◽  
In Vitro Transcription ◽  
Rna Polymerases ◽  
Hyperthermophilic Archaea ◽  
Active Centre ◽  
Α Subunit ◽  
Recent Success ◽  
Open Complex Formation

The recent success in reconstitution of RNAPs (RNA polymerases) from hyperthermophilic archaea from bacterially expressed purified subunits opens the way for detailed structure–function analyses of multisubunit RNAPs. The archaeal enzyme shows close structural similarity to eukaryotic RNAP, particularly to polymerase II, and can therefore be used as model for analyses of the eukaryotic transcriptional machinery. The cleft loops in the active centre of RNAP were deleted and modified to unravel their function in interaction with nucleic acids during transcription. The rudder, lid and fork 2 cleft loops were required for promoter-directed initiation and elongation, the rudder was essential for open complex formation. Analyses of transcripts from heteroduplex templates containing stable open complexes revealed that bubble reclosure is required for RNA displacement during elongation. Archaeal transcription systems contain, besides the orthologues of the eukaryotic transcription factors TBP (TATA-box-binding protein) and TF (transcription factor) IIB, an orthologue of the N-terminal part of the α subunit of eukaryotic TFIIE, called TFE, whose function is poorly understood. Recent analyses revealed that TFE is involved in open complex formation and, in striking contrast with eukaryotic TFIIE, is also present in elongation complexes. Recombinant archaeal RNAPs lacking specific subunits were used to investigate the functions of smaller subunits. These studies revealed that the subunits P and H, the orthologues of eukaryotic Rpb12 and Rpb5, were not required for RNAP assembly. Subunit P was essential for open complex formation, and the ΔH enzyme was greatly impaired in all assays, with the exception of promoter recruitment. Recent reconstitution studies indicate that Rpb12 and Rpb5 can be incorporated into archaeal RNAP and can complement for the function of the corresponding archaeal subunit in in vitro transcription assays.

Sign in / Sign up

Export Citation Format

Share Document