Structure of a protein L23-RNA complex located at the A-site domain of the ribosomal peptidyl transferase centre

Birte Vester, Roger Antony Garrett

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Protein L23 from the ribosome of Escherichia coli is the primary ribosomal product cross-linked to affinity-labelled puromycin; it lies, therefore, within the A-site domain of the peptidyl transferase centre on the 50 S subunit. We have characterized this functional domain by isolating and sequencing the RNA binding site of protein L23; it consists of two main fragments of 25 and 105 nucleotides that strongly interact and are separated by 172 nucleotides in the primary sequence. The higher-order structure of the RNA moiety was probed by chemical reagents, and by single-strand and double-strand-specific ribonucleases; a secondary structural model and a tertiary structural interaction are proposed on the basis of these data that are compatible with phylogenetic sequence comparisons. Several nucleotides exhibited altered chemical reactivity, both lower and higher, in the presence of protein L23, thereby implicating a large proportion of the RNA structure in the protein binding. The sites were located mainly at the extremities of the helices and at nucleotides that were putatively bulged out from the helices. The RNA moiety and an adjacent excised fragment contain several highly conserved sequences and a modified adenosine. Such sequences constitute important functional domains of the RNA and may contribute to the putative role of this RNA region in the peptidyl transferase centre.
OriginalsprogEngelsk
TidsskriftJournal of Molecular Biology
Vol/bind179
Udgave nummer3
Sider (fra-til)431-52
Antal sider22
ISSN0022-2836
StatusUdgivet - 1984

Fingeraftryk

Peptidyl Transferases
RNA
Proteins
Ribonucleases
Protein Binding

Citer dette

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title = "Structure of a protein L23-RNA complex located at the A-site domain of the ribosomal peptidyl transferase centre",
abstract = "Protein L23 from the ribosome of Escherichia coli is the primary ribosomal product cross-linked to affinity-labelled puromycin; it lies, therefore, within the A-site domain of the peptidyl transferase centre on the 50 S subunit. We have characterized this functional domain by isolating and sequencing the RNA binding site of protein L23; it consists of two main fragments of 25 and 105 nucleotides that strongly interact and are separated by 172 nucleotides in the primary sequence. The higher-order structure of the RNA moiety was probed by chemical reagents, and by single-strand and double-strand-specific ribonucleases; a secondary structural model and a tertiary structural interaction are proposed on the basis of these data that are compatible with phylogenetic sequence comparisons. Several nucleotides exhibited altered chemical reactivity, both lower and higher, in the presence of protein L23, thereby implicating a large proportion of the RNA structure in the protein binding. The sites were located mainly at the extremities of the helices and at nucleotides that were putatively bulged out from the helices. The RNA moiety and an adjacent excised fragment contain several highly conserved sequences and a modified adenosine. Such sequences constitute important functional domains of the RNA and may contribute to the putative role of this RNA region in the peptidyl transferase centre.",
keywords = "Acyltransferases, Autoradiography, Base Sequence, Binding Sites, Electrophoresis, Polyacrylamide Gel, Escherichia coli, Escherichia coli Proteins, Macromolecular Substances, Models, Molecular, Nucleic Acid Conformation, Peptidyl Transferases, Protein Biosynthesis, RNA, Ribosomal, Ribonucleases, Ribosomal Proteins, Ribosomes",
author = "Birte Vester and Garrett, {Roger Antony}",
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Structure of a protein L23-RNA complex located at the A-site domain of the ribosomal peptidyl transferase centre. / Vester, Birte; Garrett, Roger Antony.

I: Journal of Molecular Biology, Bind 179, Nr. 3, 1984, s. 431-52.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Structure of a protein L23-RNA complex located at the A-site domain of the ribosomal peptidyl transferase centre

AU - Vester, Birte

AU - Garrett, Roger Antony

PY - 1984

Y1 - 1984

N2 - Protein L23 from the ribosome of Escherichia coli is the primary ribosomal product cross-linked to affinity-labelled puromycin; it lies, therefore, within the A-site domain of the peptidyl transferase centre on the 50 S subunit. We have characterized this functional domain by isolating and sequencing the RNA binding site of protein L23; it consists of two main fragments of 25 and 105 nucleotides that strongly interact and are separated by 172 nucleotides in the primary sequence. The higher-order structure of the RNA moiety was probed by chemical reagents, and by single-strand and double-strand-specific ribonucleases; a secondary structural model and a tertiary structural interaction are proposed on the basis of these data that are compatible with phylogenetic sequence comparisons. Several nucleotides exhibited altered chemical reactivity, both lower and higher, in the presence of protein L23, thereby implicating a large proportion of the RNA structure in the protein binding. The sites were located mainly at the extremities of the helices and at nucleotides that were putatively bulged out from the helices. The RNA moiety and an adjacent excised fragment contain several highly conserved sequences and a modified adenosine. Such sequences constitute important functional domains of the RNA and may contribute to the putative role of this RNA region in the peptidyl transferase centre.

AB - Protein L23 from the ribosome of Escherichia coli is the primary ribosomal product cross-linked to affinity-labelled puromycin; it lies, therefore, within the A-site domain of the peptidyl transferase centre on the 50 S subunit. We have characterized this functional domain by isolating and sequencing the RNA binding site of protein L23; it consists of two main fragments of 25 and 105 nucleotides that strongly interact and are separated by 172 nucleotides in the primary sequence. The higher-order structure of the RNA moiety was probed by chemical reagents, and by single-strand and double-strand-specific ribonucleases; a secondary structural model and a tertiary structural interaction are proposed on the basis of these data that are compatible with phylogenetic sequence comparisons. Several nucleotides exhibited altered chemical reactivity, both lower and higher, in the presence of protein L23, thereby implicating a large proportion of the RNA structure in the protein binding. The sites were located mainly at the extremities of the helices and at nucleotides that were putatively bulged out from the helices. The RNA moiety and an adjacent excised fragment contain several highly conserved sequences and a modified adenosine. Such sequences constitute important functional domains of the RNA and may contribute to the putative role of this RNA region in the peptidyl transferase centre.

KW - Acyltransferases

KW - Autoradiography

KW - Base Sequence

KW - Binding Sites

KW - Electrophoresis, Polyacrylamide Gel

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Macromolecular Substances

KW - Models, Molecular

KW - Nucleic Acid Conformation

KW - Peptidyl Transferases

KW - Protein Biosynthesis

KW - RNA, Ribosomal

KW - Ribonucleases

KW - Ribosomal Proteins

KW - Ribosomes

M3 - Journal article

VL - 179

SP - 431

EP - 452

JO - Journal of Molecular Biology

JF - Journal of Molecular Biology

SN - 0022-2836

IS - 3

ER -