TY - JOUR
T1 - Contribution of the second OB fold of ribosomal protein S1 from Escherichia coli to the recognition of TmRNA
AU - Okada, Takahiro
AU - Wower, Iwona K.
AU - Wower, Jacek
AU - Zwieb, Christian W.
AU - Kimura, Makoto
N1 - Funding Information:
This work was supported in part by a grant from the National Project on Protein Structural and Functional Analyses, by grant-aid for scientific research from the Ministry of Education, Science, Technology, Sports and Culture of Japan, and by a grant under Rice Genome Project PR-3007, MAFF, Japan. I.K.W. and C.W.Z. were supported by National Institute of Health Grant GM-58267.
PY - 2004/11
Y1 - 2004/11
N2 - Escherichia coli ribosomal protein S1 is composed of six repeating homologous oligonucleotide/oligosaccharide-binding fold (OB folds). In trans-translation, S1 plays a role in delivering transfer-messenger RNA (tmRNA) to stalled ribosomes. The second OB fold of S1 was found to be protected from tryptic digestion in the presence of tmRNA. Truncated S1 mutant Δ2, in which the first and second OB folds were deleted, showed significantly decreased tmRNA-binding activity. Furthermore, the E. coli S1 homolog (BS1) from Bacillus subtilis, which corresponds to the four C-terminal OB folds of E. coli S1, showed no interaction with E. coli tmRNA, as judged by the results of a gel shift assay. Surface plasmon resonance analysis revealed that mutant Δ2 and BS1 had decreased association rate constants (ka, 0.59 × 10 3M-1·S-1; and ka, 1.89 × 10 3M-1·S-1), while they retained the respective dissociation rate constants (kd, 0.67 × 10-3 S -1; and kd, 0.53 × 10-3 S-1), in comparison with wild-type protein S1 (ka, 3.32 × 103 M -1·S-1; and kd, 0.56 × 10-3 S -1). These results suggest that the second OB fold in protein S1 is essential for the recognition of tmRNA, while the four C-terminal OB folds play a role in stabilizing the S1-tmRNA complex.
AB - Escherichia coli ribosomal protein S1 is composed of six repeating homologous oligonucleotide/oligosaccharide-binding fold (OB folds). In trans-translation, S1 plays a role in delivering transfer-messenger RNA (tmRNA) to stalled ribosomes. The second OB fold of S1 was found to be protected from tryptic digestion in the presence of tmRNA. Truncated S1 mutant Δ2, in which the first and second OB folds were deleted, showed significantly decreased tmRNA-binding activity. Furthermore, the E. coli S1 homolog (BS1) from Bacillus subtilis, which corresponds to the four C-terminal OB folds of E. coli S1, showed no interaction with E. coli tmRNA, as judged by the results of a gel shift assay. Surface plasmon resonance analysis revealed that mutant Δ2 and BS1 had decreased association rate constants (ka, 0.59 × 10 3M-1·S-1; and ka, 1.89 × 10 3M-1·S-1), while they retained the respective dissociation rate constants (kd, 0.67 × 10-3 S -1; and kd, 0.53 × 10-3 S-1), in comparison with wild-type protein S1 (ka, 3.32 × 103 M -1·S-1; and kd, 0.56 × 10-3 S -1). These results suggest that the second OB fold in protein S1 is essential for the recognition of tmRNA, while the four C-terminal OB folds play a role in stabilizing the S1-tmRNA complex.
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U2 - 10.1271/bbb.68.2319
DO - 10.1271/bbb.68.2319
M3 - Article
C2 - 15564671
AN - SCOPUS:11144344469
SN - 0916-8451
VL - 68
SP - 2319
EP - 2325
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
IS - 11
ER -