The donor substrate site within the peptidyl transferase loop of 23 S rRNA and its putative interactions with the CCA-end of N-blocked aminoacyl-tRNA(Phe)

Journal of Molecular Biology
B T PorseR A Garrett

Abstract

An RNA region associated with the donor substrate site, located at the base of the peptidyl transferase loop of 23 S rRNA, was subjected to a comprehensive single-site mutational study. Growth phenotypes of Escherichia coli cells were characterized on induction of synthesis of the mutated rRNAs and the mutated ribosomes were tested, selectively, for their capacity to generate peptide bonds under the conditions of the "fragment" assay. Most of the mutants exhibited dominant or recessive lethal growth phenotypes and, in general, defective growth correlated with low activities in peptide bond formation, although exceptions were observed with normal growth and low activities, and vice versa. All these phenotypes are consistent with defects occurring in the structure of the ribosomal donor site and/or the capacity of the donor substrate to enter or leave this site. A compensating base change approach was employed to test for Watson-Crick base-pairing interactions between the -CCA end of the P-site bound tRNA(Phe) and this region of the peptidyl-transferase loop. Single nucleotide substitutions were introduced into the -CCA end of tRNA(Phe) and the ability of the 3'-terminal pentanucleotide fragments to act as donor substrates was ex...Continue Reading

Citations

May 24, 2003·Cold Spring Harbor Symposia on Quantitative Biology·D F KimR Green
May 9, 1998·Proceedings of the National Academy of Sciences of the United States of America·M BocchettaA S Mankin
Jan 5, 2002·DNA Sequence : the Journal of DNA Sequencing and Mapping·L M Keith, C L Bender
Nov 25, 2004·Environmental Microbiology·V A P Martins Dos SantosK N Timmis
Aug 15, 1998·Biological Chemistry·C M Spahn, K H Nierhaus
Feb 6, 2020·Nucleic Acids Research·Anne E d'AquinoMichael C Jewett
Jan 5, 2000·Molecular Cell·D F Kim, R Green

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