The secondary structure of the protein L1 binding region of ribosomal 23S RNA. Homologies with putative secondary structures of the L11 mRNA and of a region of mitochondrial 16S rRNA

Branlant, Christiane; Krol, Alain; Machatt, Ali; Ebel, Jean‐Pierre

doi:10.1093/nar/9.2.293

Cited by 59 publications

(44 citation statements)

References 26 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3). This strongly suggested that it may thus be the yeast counterpart of this procaryotic ribosomal protein, even though no equivalent ribosomal component had previously been identified in eucaryotic cells (7,21,33). Our immunological data with anti-Ssm1p antibodies firmly establish that Ssm1p is a structural protein of the yeast ribosome (Fig.…”

Section: Discussionsupporting

confidence: 57%

“…In E. coli, L1 binds to the 23S rRNA of the large 50S subunit (7). The binding property of L1-like proteins to rRNA has been conserved in some archaebacteria (33).…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, only a few yeast ribosomal proteins have been found to be homologous to E. coli ribosomal proteins (32,67,71,72). The E. coli L1 ribosomal protein is an RNA-binding polypeptide which belongs to the large 50S subunits and binds to the 23S rRNA (7). E. coli mutants with ribosomes lacking this protein have been isolated, showing that L1 is not essential for bacterial ribosome function and assembly (10,63).…”

mentioning

confidence: 99%

See 2 more Smart Citations

The DuplicatedSaccharomyces cerevisiaeGeneSSM1Encodes a Eucaryotic Homolog of the Eubacterial and Archaebacterial L1 Ribosomal Proteins

Petitjean

Bonneaud

Lacroute

1995

Molecular and Cellular Biology

View full text Add to dashboard Cite

A previously unknown Saccharomyces cerevisiae gene, SSM1a, was isolated by screening for high-copy-number suppressors of thermosensitive mutations in the RNA14 gene, which encodes a component from the polyadenylation complex. The SSM1a gene codes for a 217-amino-acid protein, Ssm1p, which is significantly homologous to eubacterial and archaebacterial ribosomal proteins of the L1 family. Comparison of the Ssm1p amino acid sequence with that of eucaryotic polypeptides with unknown functions reveals that Ssm1p is the prototype of a new eucaryotic protein family. Biochemical analysis shows that Ssm1p is a structural protein that forms part of the largest 60S ribosomal subunit, which does not exist in a pool of free proteins. SSM1a is duplicated. The second gene copy, SSM1b, is functional and codes for an identical and functionally interchangeable Ssm1p protein. In wild-type cells, SSM1b transcripts accumulate to twice the level of SSM1a transcripts, suggesting that SSM1b is responsible for the majority of the Ssm1p pool. Haploid cells lacking both SSM1 genes are inviable, demonstrating that, in contrast with its Escherichia coli homolog, Ssm1p is an essential ribosomal protein. Deletion of the most expressed SSM1b gene leads to a severe decrease in the level of SSM1 transcript, associated with a reduced growth rate. Polysome profile analysis suggests that the primary defect caused by the depletion in Ssm1p is at the level of translation initiation.

show abstract

Section: Discussionsupporting

confidence: 57%

“…In E. coli, L1 binds to the 23S rRNA of the large 50S subunit (7). The binding property of L1-like proteins to rRNA has been conserved in some archaebacteria (33).…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

The DuplicatedSaccharomyces cerevisiaeGeneSSM1Encodes a Eucaryotic Homolog of the Eubacterial and Archaebacterial L1 Ribosomal Proteins

Petitjean

Bonneaud

Lacroute

1995

Molecular and Cellular Biology

View full text Add to dashboard Cite

show abstract

“…The putative MjaL1-binding site was synthesized in vitro as part of a 170-nucleotide transcript encompassing positions 2176Ϫ2327 of the 23S rRNA of M. jannaschii (23S rRNA Mja). The EcoL1-binding site on 23S rRNA, which has been identified earlier [34,35], was synthesized in vitro as part of a 180-nucleotide transcript encompassing positions 2076Ϫ2239 of 23S rRNA (23S rRNA Eco ). The dissociation constants and saturation levels obtained for homologous L1-rRNA complexes from M. jannaschii and E. coli were similar to those of the heterologous complexes of MjaL1 and EcoL1 with 23S rRNA Mva .…”

Section: Interaction Of Ribosomal Proteins L1 From M Vannielii M Jmentioning

confidence: 99%

Interaction of ribosomal L1 proteins from mesophilic and thermophilic Archaea and Bacteria with specific L1‐binding sites on 23S rRNA and mRNA

Köhrer

Mayer

Neumair

et al. 1998

European Journal of Biochemistry

View full text Add to dashboard Cite

In Bacteria and Archaea (formerly Archaebacteria) ribosomal protein L1 has a dual function, as a primary rRNA-binding protein and as a translational repressor which binds to its own mRNA. The L1-binding site on the mRNA exhibits high similarity in both sequence and secondary structure to the binding site for L1 on the 23 S rRNA. A sensitive membrane-filter-binding assay has been used to examine the interactions between ribosomal L1 proteins from different archaeal and bacterial species, and 23S rRNA and mRNA fragments from Methanococcus vannielii containing the MvaL1-binding site. Under standard conditions (0°C, pH 7.5, 20 mM Mg 2ϩ , 500 mM KCl), the apparent dissociation constant Kd of the homologous MvaL1-23S rRNA complex is 5 nM, the apparent dissociation constant K d of the MvaL1-mRNA complex is 0.15 µM. L1 proteins from Escherichia coli (EcoL1) and from the thermophilic Bacterium Thermus thermophilus (TthL1), and from the thermophilic Archaea Methanococcus thermolithotrophicus (MthL1), Methanococcus jannaschii (MjaL1), and Sulfolobus solfataricus (SsoL1) were tested for their affinity to the specific L1-binding sites on the 23 S rRNA and mRNA. In general, the affinity of L1 proteins from thermophilic species to the binding sites on both 23 S rRNA and mRNA is about one order of magnitude higher than that of their mesophilic counterparts. This stronger protein-RNA interaction might make a substantial contribution to the thermal tolerance of ribosomes in thermophilic organisms.Keywords : RNA-ribosomal protein L1 interaction ; filter-binding assay; Archaea ; mesophilic and thermophilic Methanococcus.Over the last two decades, a growing number of organisms living at temperatures close to or above the boiling point of water have been discovered. With a few (bacterial) exceptions, they have been classified as Archaea [1]. The adaption of these thermophilic organisms requires, amongst a number of modifications at the molecular level, a structural response of the ribosomes to ensure both high efficiency and high accuracy of the translational machinery at high temperatures. To date, very little is known about the molecular mechanisms regulating the thermal stabilization of ribosomes. Only for the hyperthermophilic Crenarchaeon, Sulfolobus solfataricus, have the mechanisms involved in the thermal stabilization of ribosomes been investigated in some detail. The heat stability of both 50S and 30S ribosomal subunits appears to be accounted for by the intrinsic stabilization of the helical rRNA domains (by a higher number of GϩC base pairs) and by a more extensive interaction between ribosomal proteins and rRNA [2Ϫ4].Ribosomal protein L1 from mesophilic and (hyper)thermophilic organisms is particularly suitable to study the specific interaction with RNA. In Bacteria and Archaea, ribosomal protein L1 has a dual function as a primary rRNA-binding protein and as a translational repressor. The primary and secondary structure of the L1-binding region on the 23S and 28S rRNA is highly conserved in Bacteria, Archaea and Eukarya ...

show abstract

“…There are several structural similarities between the Li binding site on 23S rRNA and the mRNA region around the Lll translation initiation site (8,9). Our previous deletion analysis has identified a region of the mRNA, preceding the Lii initiation site, important for feedback inhibition of Lii and Li synthesis by Li (6).…”

mentioning

confidence: 96%

Translational regulation of the L11 ribosomal protein operon of Escherichia coli: analysis of the mRNA target site using oligonucleotide-directed mutagenesis.

Baughman¹,

Nomura²

1984

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The Lii ribosomal protein operon in Escherichia coli consists of the genes for proteins Lii and Li and is feedback regulated by the translational repressor L1. The mRNA target site for this repression is located close to the translation initiation site of the first Lii cistron. Several mutant plasmid molecules carrying altered nucleotide sequences in the Li target site were constructed by site-directed in vitro mutagenesis using synthetic oligodeoxyribonucleotides. Specifically, we examined the importance of a presumptive double-stranded stem structure that is common among Li binding sites on rRNA from a variety of organisms and in LI1 mRNA. Mutational alterations that disrupt the stem structure were found to abolish translational regulation as analyzed both in vitro and in vivo. Two of the mutations were combined so that the stem structure was restored but with a different primary nucleotide sequence. This double mutant was shown to restore the original phenotype, the ability to be translationally regulated by L1. These experiments show the importance of the stem structure, but not its primary sequence, for the interaction of Li with the mRNA and support the concept that mRNA target sites share some structural features with the corresponding ribosomal protein binding sites of rRNA. MATERIALS AND METHODS Oligodeoxyribonucleotides used for mutagenesis were prepared by the phosphotriester synthesis method (10). dT-T-A-T-T-A-C-G-G-A-A-C-T-T-G-A-G was synthesized in thislaboratory and purified by polyacrylamide gel electrophoresis. Other oligonucleotides (see Fig. 3) were synthesized by Bruce Kaplan (City of Hope Research Institute) and were purified by HPLC. Mutagenesis directed with these oligonucleotides was carried out essentially as described by Zoller and Smith (1i). DNA sequence determination of mutant plasmids was carried out by the method of Maxam-Gilbert-.(12) from the HindIII site in the EcoRI fragment used as the target DNA for mutagenesis (see Fig. 2). Standard techniques were used for construction of various plasmids and M13 phage derivatives (13,14).DNA-dependent in vitro protein synthesis was carried out as described (2, 15). Protein Li, purified previously (16), was stored at -70'C and used for the in vitro experiments as described (2,6). Measurements of relative synthesis rates of r proteins in vivo were carried out as described in the legend to 5389The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.

show abstract

The secondary structure of the protein L1 binding region of ribosomal 23S RNA. Homologies with putative secondary structures of the L11 mRNA and of a region of mitochondrial 16S rRNA

Cited by 59 publications

References 26 publications

The DuplicatedSaccharomyces cerevisiaeGeneSSM1Encodes a Eucaryotic Homolog of the Eubacterial and Archaebacterial L1 Ribosomal Proteins

The DuplicatedSaccharomyces cerevisiaeGeneSSM1Encodes a Eucaryotic Homolog of the Eubacterial and Archaebacterial L1 Ribosomal Proteins

Interaction of ribosomal L1 proteins from mesophilic and thermophilic Archaea and Bacteria with specific L1‐binding sites on 23S rRNA and mRNA

Translational regulation of the L11 ribosomal protein operon of Escherichia coli: analysis of the mRNA target site using oligonucleotide-directed mutagenesis.

Contact Info

Product

Resources

About