Bases in 16S rRNA important for subunit association, tRNA binding, and translocation

Xinying Shi, Katie Chiu, Srikanta Ghosh, Simpson Joseph

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Ribosomes are the cellular machinery responsible for protein synthesis. A well-orchestrated step in the elongation cycle of protein synthesis is the precise translocation of the tRNA-mRNA complex within the ribosome. Here we report the application of a new in vitro modification-interference method for the identification of bases in 16S rRNA that are essential for translocation. Our results suggest that conserved bases U56, U723, A1306, A1319, and A1468 in 16S rRNA are important for translocation. These five bases were deleted or mutated so their role in translation could be studied. Depending on the type of mutation, we observed inhibition of growth rate, subunit association, tRNA binding, and/or translocation. Interestingly, deletion of U56 or A1319 or mutation of A1319 to C showed a lethal phenotype and were defective in protein synthesis in vitro. Further analysis showed that deletion of U56 or A1319 caused defects in 30S subunit assembly, subunit association, and tRNA binding. In contrast, the A1319C mutation showed no defects in subunit association; however, the extent of tRNA binding and translocation was significantly reduced. These results show that conserved bases located as far as 100 Å from the tRNA binding sites can be important for translation.

Original languageEnglish (US)
Pages (from-to)6772-6782
Number of pages11
JournalBiochemistry
Volume48
Issue number29
DOIs
StatePublished - Jul 28 2009
Externally publishedYes

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Transfer RNA
Association reactions
Ribosomes
Mutation
Defects
Proteins
Machinery
Elongation
Binding Sites
Phenotype
Messenger RNA
Growth
In Vitro Techniques

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bases in 16S rRNA important for subunit association, tRNA binding, and translocation. / Shi, Xinying; Chiu, Katie; Ghosh, Srikanta; Joseph, Simpson.

In: Biochemistry, Vol. 48, No. 29, 28.07.2009, p. 6772-6782.

Research output: Contribution to journalArticle

Shi, Xinying ; Chiu, Katie ; Ghosh, Srikanta ; Joseph, Simpson. / Bases in 16S rRNA important for subunit association, tRNA binding, and translocation. In: Biochemistry. 2009 ; Vol. 48, No. 29. pp. 6772-6782.
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