| 1. |
- Kirsebom, Leif A, et al.
(författare)
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Effects of growth conditions and mutations in RNA polymerase on translational activity in vitro in Escherichia coli
- 1980
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Ingår i: Molecular General Genetics. - 0026-8925 .- 1432-1874. ; 180:1, s. 27-33
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Tidskriftsartikel (refereegranskat)abstract
- The translational capacity in vitro in Escherichia coli, using RNA from phage R17 or Q beta as messenger, is several times higher if the extracts are prepared from cells harvested in early exponential phase or grown under conditions of good aeration compared to if extracts are prepared from cells harvested in a later growth phase or grown under semi-aerobic conditions. In low activity extracts the production of phage replicase protein is preferentially affected. Growth of a wild type strain under semi-aerobic conditions has a less pronounced effect on translational capacity in vitro using crude mRNA from normal or T4 infected cells or with poly(U). Mutants were fortuitously found which did not show a lowered translational activity in vitro as a result of entering late phase of growth. Two of these were changed in RNA polymerase. Two different translational inhibitors can be demonstrated in the ribosomal wash fraction obtained from semi-aerobically grown wild type cells, whereas only one was found in the case of aerobically grown cells. The low translational activity of semi-aerobically grown cells in vitro is implied to be dependent on the induction or activation of a translational inhibitor. It behaves like a protein but is not likely to be a protease or RNAse.
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| 2. |
- Kirsebom, Leif A, et al.
(författare)
-
Functional interactions in vivo between suppressor tRNA and mutationally altered ribosomal protein S4
- 1986
-
Ingår i: Molecular General Genetics. - 0026-8925 .- 1432-1874. ; 205:2, s. 240-247
-
Tidskriftsartikel (refereegranskat)abstract
- Ribosomal mutants (rpsD) which are associated with a generally increased translational ambiguity were investigated for their effects in vivo on individual tRNA species using suppressor tRNAs as models. It was found that nonsense suppression is either increased, unaffected or decreased depending on the codon context and the rpsD allele involved as well as the nature of the suppressor tRNA. Missense suppression of AGA and AGG by glyT(SuAGA/G) tRNA as well as UGG by glyT(SuUGG-8) tRNA is unaffected whereas suppression of UGG by glyT(SuUGA/G) or glyV(SuUGA/G) tRNA is decreased in the presence of an rpsD mutation. The effects on suppressor tRNA are thus not correlated with the ribosomal ambiguity (Ram) phenotype of the rpsD mutants used in this study. It is suggested that the mutationally altered ribosomes are changed in functional interactions with the suppressor tRNA itself rather than with the competing translational release factor(s) or cognate aminoacyl tRNA. The structure of suppressor tRNA, particularly the anticodon loop, and the suppressed codon as well as the codon context determine the allele specific functional interactions with these ribosomal mutations.
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