12.5
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0 12.5 250 25 50 100 U G C A 500 1000 2500 5000 0 12.5 250 25 50 100 500 1000 2500 5000 Trp (μM) Ribosomes at Pro- 24 Ribosomes at Trp- 12 because of lack of Trp in the AA mix Wt A2058G Ribosomes at Pro- 24 Ribosomes at Trp- 12 because of lack of Trp in the AA mix U2609C ribosomes 0 12.5 100 1000 5000 Toe-printing, accumulation of stalled ribosomes in Trp (μM)
description
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. - PowerPoint PPT Presentation
Transcript of 12.5
0
12.5 250
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100U GC A 500
1000
2500
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12.5 250
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5000Trp (μM)
Ribosomes at Pro-24
Ribosomes at Trp-12 because of lack of Trp in the AA mix
Wt A2058G
Ribosomes at Pro-24
Ribosomes at Trp-12 because of lack of Trp in the AA mix
U2609C ribosomes
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Toe-printing, accumulation of stalled ribosomes in vitro
Trp (μM)
TnaC- tRNAPro
TnaC- tRNAPro
TnaC
Wild Type A2058G
A2058T
0 0.2 0.5 1 2 5 10 20L-Trp (mM)
TnaC- tRNAPro
TnaC
0 0.2 0.5 1 2 5 10 20L-Trp (mM)
0 0.2 0.5 1 2 5 10 20L-Trp (mM)
L-Trp (mM)
Tna
C-t
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AP
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Accumulation of TnaC-tRNAPro in vitro
Be
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MU
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1-M-Trp (mM)
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23S rRNA wt
23S rRNA wt/W12R
23S rRNA A2058T
23S rRNA A2058G
Expression of tnaA’-’lacZ reporter gene in vivo
Conclusion
• The ribosomes containing the A2058G mutant 23S rRNA are less sensitive to Trp.
• The change A2058G might affect the Trp binding site.
