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U2609C ribosomes. Wt. A2058G. 12.5. 50. 250. 1000. 5000. 12.5. 50. 250. 1000. 5000. 12.5. 1000. Trp (μM). Trp (μM). 0. 100. 5000. 0. 25. 100. 500. 2500. 0. 25. 100. 500. 2500. C. U. A. G. Ribosomes at Trp-12 because of lack of Trp in the AA mix.

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12.5

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  1. U2609C ribosomes Wt A2058G 12.5 50 250 1000 5000 12.5 50 250 1000 5000 12.5 1000 Trp (μM) Trp (μM) 0 100 5000 0 25 100 500 2500 0 25 100 500 2500 C U A G Ribosomes at Trp-12 because of lack of Trp in the AA mix Ribosomes at Trp-12 because of lack of Trp in the AA mix Ribosomes at Pro-24 Ribosomes at Pro-24 Toe-printing, accumulation of stalled ribosomes in vitro

  2. Wild Type A2058G 0 0.2 0.5 1 2 5 10 20 L-Trp(mM) 0 0.2 0.5 1 2 5 10 20 L-Trp(mM) TnaC- tRNAPro TnaC- tRNAPro TnaC  TnaC A2058T 0 0.2 0.5 1 2 5 10 20 L-Trp(mM) TnaC- tRNAPro TnaC-tRNAPro accumulation L-Trp(mM) Accumulation of TnaC-tRNAPro in vitro

  3. Beta-Gal expression (MU) 1-M-Trp (mM) Expression of tnaA’-’lacZreporter gene in vivo

  4. Conclusion • The ribosomes containing the A2058G mutant 23S rRNA are less sensitive to Trp. • The change A2058G might affect the Trp binding site.

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