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Development of Genetic System for Isolation of SSU rRNA Mutants that Bypass SecM-Mediated Ribosome Stalling  

Ha, Hye-Jeong (Department of Life Science, Chung-Ang University)
Kim, Hong-Man (Department of Life Science, Chung-Ang University)
Yeom, Ji-Hyun (Department of Life Science, Chung-Ang University)
Lee, Kang-Seok (Department of Life Science, Chung-Ang University)
Publication Information
Korean Journal of Microbiology / v.44, no.4, 2008 , pp. 271-276 More about this Journal
Abstract
Ribosome stalling by nascent sticky peptide has been reported in several organisms across the kingdom. To test whether small subunit (SSU) rRNA is involved in this phenomenon, we developed a genetic system that utilized the specialized ribosome system to isolate SSU rRNA mutants that enable ribosomes to bypass the SecM-derived sticky peptide in protein synthesis. In this system, CAT-SecM mRNA, which encodes CAT protein containing the sticky peptide derived from SecM, is only translated by specialized ribosomes. These ribosomes were shown to transiently stall on CAT-SecM mRNA followed by the synthesis of the sticky peptide. Expression of specialized ribosomes resulted in the decreased steady-state level of CAT-SecM mRNA, which is consistent with a notion that ribosome stalling induces mRNA degradation. Isolation and characterization of SSU rRNA mutations using this genetic system that are sufficient to circumvent ribosome stalling induced by the SecM-derived sticky peptide will provide evidence of SSU rRNA function in mRNA cleavage.
Keywords
protein synthesis; ribosome stalling; SecM; SSU rRNA; sticky peptide;
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