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KMID : 0364820080440040271
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Korean Journal of Microbiology
2008 Volume.44 No. 4 p.271 ~ p.276
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Development of Genetic System for Isolation of SSU rRNA Mutants that Bypass SecMMediated Ribosome Stalling
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Ha Hye-Jeong
Kim Hong-Man
Yeom Ji-Hyun
Lee Kang-Seok
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Abstract
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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 SecMderived 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.
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KEYWORD
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protein synthesis, ribosome stalling, SecM, SSU rRNA, sticky peptide
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