The Folding of RNA in the 30S Subunit of Escherichia coli Ribosomes

Spitnik‐Elson, Pnina; Elson, David

doi:10.1007/978-1-4612-4884-2_13

Cited by 2 publications

(1 citation statement)

References 39 publications

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“…Apparently, a long range interaction exists between the base-paired 1192 region and the 1410-1490 stem structure. To our knowledge, no such interaction involving these areas has been reported (see also Spitnik-Elson and Elson, 1985).…”

Section: Discussionmentioning

confidence: 84%

Mutagenesis at the mRNA decoding site in the 16S ribosomal RNA using the specialized ribosome system in Escherichia coli.

Hui¹,

Eaton²,

Boer³

1988

The EMBO Journal

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In the specialized ribosome system, a distinct pool of mutated ribosomes is dedicated to the translation of one particular mRNA species. This was accomplished by altering the Shine-LDalgarno sequence on the mRNA and its complementary anti-Shine -Dalgarno sequence on the plasmid-borne 16S rRNA gene. Here, using the specialized ribosome system, we were able to introduce mutations in key regions of the 16S rRNA and could study their effect on translation in vivo. The C 1400 region has been implicated to play a role in the actual mRNA decoding process. Several ribosomal mutations were introduced in this region. We showed that substitution of the evolutionary highly conserved C1400 residue by a G-or an A-residue inhibits ribosomal activity by 80% and 50% respectively, whereas, a C to a U change at this conserved position does not affect overall ribosomal activity. The adjacent stem structure (1410-1490) was also examined. Disruption of the stem by replacing either one of the arms of this stem, with a different sequence, inhibits ribosomal activity by -80%. A small but significant restoration of translation could be achieved by recreating a complementary stem with a different sequence. We found that full reversion of activity could be obtained when such mutated ribosomes were made spectinomycin resistant by introducing a C to A substitution at position 1192 which is located far away in the secondary structure map of the 16S rRNA molecule. Based on these results we conclude that some, but not all, of the nucleotides in the conserved C1400 region play a key role in translation. Most importantly, we show that the specialized ribosome system provides a very powerful tool for structure and function studies of any region in the complex ribosomal RNA molecule.

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Section: Discussionmentioning

confidence: 84%

Mutagenesis at the mRNA decoding site in the 16S ribosomal RNA using the specialized ribosome system in Escherichia coli.

Hui¹,

Eaton²,

Boer³

1988

The EMBO Journal

View full text Add to dashboard Cite

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Conformational analysis of 16 S ribosomal RNA from Escherichia coli by scanning transmission electron microscopy

et al. 1988

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Digitized images of molecules of 16 S rRNA from Escherichiu coli, obtained by scanning transmission electron microscopy (STEM), provide quantitative structural information that is lacking in conventional electron micrographs. We have determined the morphology, total molecular mass, mass distribution within individual rRNA molecules and apparent radii of gyration. From the linear density (M/L) we have assessed the number of strands in the structural backbone of rRNA and studied the pattern of branching and folding related to the secondary and tertiary structure of rRNAs under various buffer conditions. Even in reconstitution buffer 16 S RNA did not show any resemblance to the native 30 S subunit.rRNA; Electron microscopy; RNA structure

show abstract

The Folding of RNA in the 30S Subunit of Escherichia coli Ribosomes

Cited by 2 publications

References 39 publications

Mutagenesis at the mRNA decoding site in the 16S ribosomal RNA using the specialized ribosome system in Escherichia coli.

Mutagenesis at the mRNA decoding site in the 16S ribosomal RNA using the specialized ribosome system in Escherichia coli.

Conformational analysis of 16 S ribosomal RNA from Escherichia coli by scanning transmission electron microscopy

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