Kinetic analysis of ribosome binding process onto mRNA using a quartz-crystal microbalance - RNA

Kinetic analysis of ribosome binding process onto mRNA using a quartz-crystal microbalance
2006
Shuntaro Takahashi1, Ryoko Akita1, Hiroyuki Furusawa1, Yoshihiro Shimizu2, Takuya Ueda2 and Yoshio Okahata1
1 Department of Biomolecular Engineering, Frontier Collaborative Research Center, Tokyo Institute of Technology and CREST, Japan Science and Technology Corporation (JST), 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan, 2 Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, FSB401, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan
Nucleic Acids Symposium Series 2006
Translation initiation is the most dynamic and important step along a series of protein synthesis processes. In bacteria, it is generally accepted that the 70S ribosome initially dissociates into the 30S and 50S subunit, and then, the 30S ribosomal subunit binds to the Shine-Dargalno (SD) sequence of mRNA. We analyzed binding kinetics of 70S, 50S and 30S ribosomes to the SD sequence by using a mRNA-immobilized 27 MHz quartz-crystal microbalance (QCM). The 70S ribosome was found to bind strongly to the SD sequence as a ratio of 1:1 without dissociation to each subunit from the lateral side, as well as the 30S subunit. The binding constant for 70S increased in the presence of the initiator tRNA, which suggests that the SD and initiator codon of AUG could be also recognized precisely with 70S.
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