Role of 5S RNA in assembly and function of the 50S subunit from Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 73 ·No. 7 ·1976-07-00 ·Pages 2221-5

Dohme F, Nierhaus KH

Abstract

Total reconstitution experiments performed under various conditions revealed that 5S RNA plays an important role during the last assembly step in vitro leading to an active 50S particle. For the preceding steps this RNA species is dispensable. However, 50S RNA can be integrated efficiently during any of the assembly steps in vitro. The 47S particle, reconstituted in two steps and lacking 5S RNA, shows low but significant activity in many functional tests. High activity could be obtained by incubating this particle with 5S RNA alone, demonstrating the importance of the 5S RNA in generating an active ribosomal conformation. In particular, the activity of the peptidyltransferase (peptidyl-tRNA:aminoacyl-tRNA N-peptidyltransferase; EC 2.3.2.12) center is drastically influenced by 5S RNA. No significant factor-dependent tRNA binding to the A-site was observed with the 47S particle, in contrast to the corresponding P-site binding. The elongation factor G dependent GTPase activity was not affected by the lack of 5S RNA.

MeSH Terms

Binding Sites Cell-Free System Escherichia coli GTP Phosphohydrolase-Linked Elongation Factors/metabolism Peptide Elongation Factors Peptidyl Transferases RNA, Bacterial/metabolism,physiology RNA, Ribosomal/metabolism,physiology RNA, Transfer/metabolism Ribosomal Proteins/metabolism Ribosomes/metabolism,ultrastructure Structure-Activity Relationship

Chemicals

Peptide Elongation Factors RNA, Bacterial RNA, Ribosomal Ribosomal Proteins RNA, Transfer Peptidyl Transferases GTP Phosphohydrolase-Linked Elongation Factors

Authors & Affiliations

2 authors, click to expand affiliations / ORCID

Dohme F

Nierhaus K H

References (23)

23 references, click to expand

Substrate and antibiotic binding sites at the peptidyl transferase centre of E. coli ribosomes.
FEBS Lett. 1970 Feb 16;6(3):273-277 PMID: 11947393
Nucleotide sequence of KB cell 5S RNA.
Science. 1967 Dec 29;158(3809):1695-9 PMID: 6059650
Identification of the chloramphenicol-binding protein in Escherichia coli ribosomes by partial reconstitution.
Proc Natl Acad Sci U S A. 1973 Aug;70(8):2224-8 PMID: 4365366
A protein involved in the peptidyltransferase activity of Escherichia coli ribosomes.
Proc Natl Acad Sci U S A. 1973 Jul;70(7):1931-5 PMID: 4579005
Ribosomal proteins. 43. In vivo assembly of Escherichia coli ribosomal proteins.
J Mol Biol. 1973 Mar 15;74(4):587-97 PMID: 4580907
Studies on the formation of transfer ribonucleic acid-ribosome complexes. IV. A new assay for codon recognition and interaction of transfer ribonucleic acid with 50 S subunits.
J Biol Chem. 1968 Aug 10;243(15):4038-44 PMID: 4875320
Ribosomal proteins. XXII. Studies on the altered protein S5 from a spectinomycin-resistant mutant of Escherichia coli.
J Mol Biol. 1972 Feb 28;64(1):201-9 PMID: 4259369
The involvement of 5S RNA in the binding of tRNA to ribosomes.
Biochem Biophys Res Commun. 1973 Oct 1;54(3):942-8 PMID: 4584885
ATPase and GTPase activities associated with the 5-S RNA-protein complex of Escherichia coli ribosomes.
Biochim Biophys Acta. 1975 May 1;390(2):226-30 PMID: 239743
Total reconstitution of functionally active 50S ribosomal subunits from Escherichia coli.
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4713-7 PMID: 4612527
Biosynthesis of ribosomes in E. coli. I. Properties of ribosomal precursor particles and their RNA components.
Biochimie. 1971;53(3):369-82 PMID: 4935405
The association of ribosomal subunits of Escherichia coli. 1. Two types of association products differing in their apparent sedimentation coefficient.
Eur J Biochem. 1973 Nov 15;39(2):499-510 PMID: 4590451
Role of 5S RNA in the functions of 50S ribosomal subunits.
Proc Natl Acad Sci U S A. 1971 Dec;68(12):2932-6 PMID: 5289238
Kirromycin, an inhibitor of protein biosynthesis that acts on elongation factor Tu.
Proc Natl Acad Sci U S A. 1974 Dec;71(12):4910-4 PMID: 4373734
Function of sulfhydryl groups in ribosome-elongation factor G reactions. Assignment of guanine nucleotide binding site to elongation factor G.
J Biol Chem. 1975 Nov 10;250(21):8344-52 PMID: 172495
Structural and functional studies of ribonucleoprotein fragments isolated from Escherichia coli 50 S ribosomal subunits.
J Mol Biol. 1975 May 5;94(1):111-21 PMID: 124783
Role of 5S ribosomal RNA in polypeptide synthesis.
Biochem Biophys Res Commun. 1969 Aug 22;36(5):711-20 PMID: 4980101
Tables for estimating sedimentation through linear concentration gradients of sucrose solution.
Anal Biochem. 1967 Jul;20(1):114-49 PMID: 6034985
Studies on the attachment and release of 5 s ribosomal RNA from the large ribosomal subunit.
J Mol Biol. 1969 Jan 14;39(1):31-44 PMID: 4999871
Binding of aminoacyl tRNA to ribosomes: a function of 5S-RNA.
Acta Biol Med Ger. 1974;33(5-6):605-8 PMID: 4469374
Ribosomal proteins. VII. Two-dimensional polyacrylamide gel electrophoresis for fingerprinting of ribosomal proteins.
Anal Biochem. 1970 Aug;36(2):401-12 PMID: 4916449
ATPase and GTPase activities associated with a specific 5S RNA-protein complex.
Proc Natl Acad Sci U S A. 1973 Oct;70(10):2870-3 PMID: 4355369
Protein involved in the binding of dihydrostreptomycin to ribosomes of Escherichia coli.
J Mol Biol. 1973 Nov 25;81(1):71-82 PMID: 4129206

Article Info

Journal

Proceedings of the National Academy of Sciences of the United States of America

Abbr.

Proc Natl Acad Sci U S A

ISSN

0027-8424

DOI

10.1073/pnas.73.7.2221

Published

1976-07-00

Pages

2221-5

Language

English

Region

United States

NLM ID

7505876

PMCID

PMC430504

Subset

External Links

PubMed source DOI: 10.1073/pnas.73.7.2221 PMC Full Text View journal details

Analysis Services

Role of 5S RNA in assembly and function of the 50S subunit from Escherichia coli.

Contact

WeChat Official Account