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. 1990 May;87(10):3700–3704. doi: 10.1073/pnas.87.10.3700

Base changes at position 792 of Escherichia coli 16S rRNA affect assembly of 70S ribosomes.

M Santer 1, E Bennett-Guerrero 1, S Byahatti 1, S Czarnecki 1, D O'Connell 1, M Meyer 1, J Khoury 1, X Cheng 1, I Schwartz 1, J McLaughlin 1
PMCID: PMC53970  PMID: 2140191

Abstract

To investigate the function of base 792 of 16S rRNA in 30S ribosomes of Escherichia coli, the wild-type (adenine) residue was changed to guanine, cytosine, or uracil by oligonucleotide-directed mutagenesis. Each base change conferred a unique phenotype on the cells. Cells containing plasmid pKK3535 with G792 or T792 showed no difference in generation time in LB broth containing ampicillin, whereas cells with C792 exhibited a 20% increase in generation time in this medium. To study the effect on cell growth of a homogeneous population of mutant ribosomes, the mutations were cloned into the 16S rRNA gene on pKK3535 carrying a spectinomycin-resistance marker (thymine at position 1192), and the cells were grown with spectinomycin. Cells containing G792 or C792 showed 16% and 56% increases in generation time, respectively, and a concomitant decrease in 35S assimilation into proteins. Cells with T792 did not grow in spectinomycin-containing medium. Maxicell analyses indicated decreasing ability to form 70S ribosomes from 30S subunits containing guanine, cytosine, or uracil at position 792 in 16S rRNA. It appeared that C792-containing 30S ribosomes had lost the ability to bind initiation factor 3.

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Selected References

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