PMID:10214965
| Citation |
Fujiwara, T, Ito, K, Nakayashiki, T and Nakamura, Y (1999) Amber mutations in ribosome recycling factors of Escherichia coli and Thermus thermophilus: evidence for C-terminal modulator element. FEBS Lett. 447:297-302 |
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| Abstract |
Ribosome recycling factor, referred to as RRF, is essential for bacterial growth because of its activity of decomposition of the post-termination complex of the ribosome after release of polypeptides. In this study, we isolated a conditionally lethal amber mutation, named frr-3, in the Escherichia coli RRF gene at amino acid position 161, showing that the truncation of the C-terminal 25 amino acids of RRF is lethal to E. coli. An RRF gene cloned from Thermus thermophilus, whose protein is 44% identical and 68% similar to E. coli RRF, failed to complement the frr-3(Am) allele. However, truncation of the C-terminal five amino acids conferred intergeneric complementation activity on T. thermophilus RRF, demonstrating the modulator activity of the C-terminal tail. Rapid purification of T. thermophilus RRF was achieved by T7-RNA polymerase-driven overexpression for crystallography. |
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| Keywords |
Amino Acid Sequence; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Base Sequence; Cloning, Molecular; DNA Primers/genetics; DNA, Bacterial/genetics; Escherichia coli/genetics; Escherichia coli/metabolism; Genes, Bacterial; Genetic Complementation Test; Molecular Sequence Data; Mutation; Proteins/genetics; Proteins/metabolism; Ribosomal Proteins; Ribosomes/metabolism; Species Specificity; Thermus thermophilus/genetics; Thermus thermophilus/metabolism |
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