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Nucleic Acids Symposium Series 2007 51(1):45-46; doi:10.1093/nass/nrm023
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© 2007 Oxford University Press

Kinetic analysis of the effects of translation enhancers in translation initiation

Shuntaro Takahashi1, Hiroyuki Furusawa1, Yoshihiro Shimizu2, Takuya Ueda2 and Yoshio Okahata1,*

1Department of Biomolecular Engineering, Frontier Collaborative Research Center, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan and
2Department of Medical Genome Sciences, Graduate School of Frontier Sciences, University of Tokyo, FSB401, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan.

*Corresponding author. E-mail: yokahata{at}bio.titech.ac.jp

Abstract

Translation initiation is the most important step within a series of protein biosynthesis processes because the incorporation of ribosomes to a mRNA mainly determines efficiencies of translation. In bacteria, translation enhancers located on the 5’ upstream of the Shine-Dargalno (SD) sequence on mRNAs are known to accelerate the efficiency of protein biosynthesis. To investigate the role of translation enhancers in translation initiation, we analyzed binding kinetics of a 30S ribosomal subunit to a mRNA immobilized on a 27 MHz quartz-crystal microbalance (QCM). The association constant (Ka) was rather low for the mRNA including a translation enhancer sequence compared with that for the mRNA without translation enhancers. These kinetic parameters suggest that translation enhancers destabilize the ribosome-mRNA complex on an SD sequence to move on the next step of decoding its mRNA.


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