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. 2001 May;2(5):399-402.
doi: 10.1093/embo-reports/kve091.

Ribosome-associated protein that inhibits translation at the aminoacyl-tRNA binding stage

D E Agafonov  1 , V A Kolb, A S Spirin
Affiliations

Ribosome-associated protein that inhibits translation at the aminoacyl-tRNA binding stage

D E Agafonov et al. EMBO Rep. 2001 May.

Abstract

We have recently isolated and characterized a novel protein associated with Escherichia coli ribosomes and named protein Y (pY). Here we show that the ribosomes from bacterial cells growing at a normal physiological temperature contain no pY, whereas a temperature downshift results in the appearance of the protein in ribosomes. The protein also appears in the ribosomes of those cells that reached the stationary phase of growth at a physiological temperature. Our experiments with cell-free translation systems demonstrate that the protein inhibits translation at the elongation stage by blocking the binding of aminoacyl-tRNA to the ribosomal A site. The function of the protein in adaptation of cells to environmental stress is discussed.

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Figures

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Fig. 1. Cold-induced synthesis of pY in E. coli cells. Shown are fragments of gel slabs stained with Coomassie Blue G 250. (A) Proteins from ribosomes of cells grown at 37°C. (B) Proteins from ribosomes of cells subjected to a temperature downshift.
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Fig. 2. Effect of pY on cell-free translation of GFP mRNA. Molar ratio of ribosomes to pY is indicated above bars. The amount of newly synthesized protein was determined from the radioactivity of hot 10% trichloroacetic acid precipitate.
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Fig. 3. Inhibition of poly(U)-directed polyphenylalanine elongation by pY. The molar ratio of ribosomes to pY is indicated above bars.
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Fig. 4. Blocking of aminoacyl-tRNA binding to the ribosomal A site by pY. (A) Binding of [14C]phenylalanyl-tRNAPhe–EF-Tu–GTP ternary complex to 70S ribosomes. (B) Binding of [14C]phenylalanyl-tRNAPhe to 70S ribosomes.
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Fig. 5. Binding of [3H]tetracycline to 70S ribosomes in the presence of pY. The molar ratio of ribosomes to pY is indicated above bars.
See this image and copyright information in PMC

References

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