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).

B T Porse; H P Thi-Ngoc; R A Garrett

doi:10.1006/jmbi.1996.0655

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).

B T Porse, H P Thi-Ngoc, R A Garrett

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

37 Citationer (Scopus)

Abstract

Udgivelsesdato: 1996-Dec-6

Originalsprog	Engelsk
Tidsskrift	Journal of Molecular Biology
Vol/bind	264
Udgave nummer	3
Sider (fra-til)	472-83
Antal sider	11
ISSN	0022-2836
DOI	https://doi.org/10.1006/jmbi.1996.0655
Status	Udgivet - 1996

Bibliografisk note

Keywords: Escherichia coli; Peptide Chain Elongation, Translational; Peptidyl Transferases; Point Mutation; RNA, Ribosomal, 23S; RNA, Transfer, Amino Acyl; Ribonucleosides; Ribosomes

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10.1006/jmbi.1996.0655

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Citationsformater

@article{75b0888058a311dd8d9f000ea68e967b,

title = "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).",

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 examined for ribosomes carrying the different mutated 23 S rRNAs. No evidence was found for the occurrence of Watson-Crick base-pairing interactions. However, the data are consistent with the formation of a Hoogsteen pair between the 3'-terminal adenosine base of the donor substrate and U2585 of the 23 S rRNA.",

author = "Porse, {B T} and Thi-Ngoc, {H P} and Garrett, {R A}",

note = "Keywords: Escherichia coli; Peptide Chain Elongation, Translational; Peptidyl Transferases; Point Mutation; RNA, Ribosomal, 23S; RNA, Transfer, Amino Acyl; Ribonucleosides; Ribosomes",

year = "1996",

doi = "10.1006/jmbi.1996.0655",

language = "English",

volume = "264",

pages = "472--83",

journal = "Journal of Molecular Biology",

issn = "0022-2836",

publisher = "Academic Press",

number = "3",

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T1 - 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).

AU - Porse, B T

AU - Thi-Ngoc, H P

AU - Garrett, R A

N1 - Keywords: Escherichia coli; Peptide Chain Elongation, Translational; Peptidyl Transferases; Point Mutation; RNA, Ribosomal, 23S; RNA, Transfer, Amino Acyl; Ribonucleosides; Ribosomes

PY - 1996

Y1 - 1996

N2 - 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 examined for ribosomes carrying the different mutated 23 S rRNAs. No evidence was found for the occurrence of Watson-Crick base-pairing interactions. However, the data are consistent with the formation of a Hoogsteen pair between the 3'-terminal adenosine base of the donor substrate and U2585 of the 23 S rRNA.

AB - 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 examined for ribosomes carrying the different mutated 23 S rRNAs. No evidence was found for the occurrence of Watson-Crick base-pairing interactions. However, the data are consistent with the formation of a Hoogsteen pair between the 3'-terminal adenosine base of the donor substrate and U2585 of the 23 S rRNA.

U2 - 10.1006/jmbi.1996.0655

DO - 10.1006/jmbi.1996.0655

M3 - Journal article

C2 - 8969299

SN - 0022-2836

VL - 264

SP - 472

EP - 483

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

IS - 3

ER -