Polyamines Accelerate Codon Recognition by Transfer RNAs on the Ribosome

被引:18
|
作者
Hetrick, Byron [1 ]
Khade, Prashant K. [1 ]
Lee, Kristin [1 ]
Stephen, Jenise [1 ]
Thomas, Alex [1 ]
Joseph, Simpson [1 ]
机构
[1] Univ Calif San Diego, Dept Chem & Biochem, La Jolla, CA 92093 USA
基金
美国国家卫生研究院;
关键词
AMINOACYL-TRANSFER-RNA; ELONGATION-FACTOR TU; ESCHERICHIA-COLI RIBOSOMES; MESSENGER-RNA; PROTEIN-BIOSYNTHESIS; ACID INCORPORATION; TERNARY COMPLEXES; CRYSTAL-STRUCTURE; P-SITE; SELECTION;
D O I
10.1021/bi1009776
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The selection of aminoacyl-tRNAs by the ribosome is a fundamental step in the elongation cycle of protein synthesis. tRNA selection is a multistep process that ensures only correct aminoacyl-tRNAs are accepted while incorrect aminoacyl-tRNAs are rejected. A key step in tRNA selection is the formation of base pairs between the anticodon of the aminoacyl-tRNA and the m RNA codon in the A site; called "codon recognition". Here, we report the development of a new, fluorescence-based, kinetic assay for monitoring codon recognition by the ribosome. Using this assay, we show that codon recognition is a second-order binding step under optimal conditions. Additionally, we show that at low Mg2+ concentrations, the polyamines spermine and spermidine stimulate codon recognition by the ribosome without a loss of fidelity. Polyamines may accelerate codon recognition by altering the structure and dynamics of the anticodon arm of the aminoacyl-tRNA.
引用
收藏
页码:7179 / 7189
页数:11
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