The L7/L12 ribosomal domain of the ribosome: structural and functional studies

Gudkov, A.T.

doi:10.1016/s0014-5793(97)00361-x

Cited by 59 publications

(41 citation statements)

References 57 publications

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“…They have been described to target and bind to elongation factor 2 1,2 (18), but it was also immediately apparent that the inhibitor's mode of action involved additional cellular components, because resistant mutants were obtained outside EF2 1 , and high affinity binding of the drug to macromolecules required the presence of ribosomes, 2 suggesting that the binding site on EF2 is fully formed only after the factor interacts with a ribosome. In this work, experiments are described showing that rpP0, an essential component of the ribosomal large subunit stalk, plays a large role in determining the sensitivity to sordarin antifungal GM193663, as single point mutations in rpP0, and substitution of heterologous rpP0 proteins for the endogenous one, reduce measurably the sensitivity of yeast toward these compounds (Tables II and III).…”

Section: Discussionmentioning

confidence: 99%

“…Sordarin inhibitors seem to bind to elongation factor 2 1,2 (18), and point mutations on the factor make cells resistant to the inhibitors 1 (18), but high affinity binding requires the presence of ribosomes, 2 and resistant mutants were detected, which did not map on EF2. 1 This work describes the gene mutated in a second complementation group of S. cerevisiae mutants resistant to GM193663, a potent sordarin derivative.…”

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confidence: 99%

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Ribosomal P-protein Stalk Function Is Targeted by Sordarin Antifungals

Gómez-Lorenzo¹,

Garcia-Bustos²

1998

Journal of Biological Chemistry

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Sordarin derivatives are remarkably selective inhibitors of fungal protein synthesis. Available evidence points to a binding site for these inhibitors on elongation factor 2, but high affinity binding requires the presence of ribosomes. The gene mutated in one of the two isolated complementation groups of Saccharomyces cerevisiae mutants resistant to the sordarin derivative GM193663 has now been identified. It is RPP0, encoding the essential protein of the large ribosomal subunit stalk rpP0. Resistant mutants are found to retain most of the binding capacity for the drug, indicating that mutations in rpP0 endow the ribosome with the capacity to perform translation elongation in the presence of the inhibitor. Other proteins of the ribosomal stalk influence the expression of resistance, pointing to a wealth of interactions between stalk components and elongation factors. The involvement of multiple elements of the translation machinery in the mode of action of sordarin antifungals may explain the large selectivity of these compounds, even though the individual target components are highly conserved proteins.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Ribosomal P-protein Stalk Function Is Targeted by Sordarin Antifungals

Gómez-Lorenzo¹,

Garcia-Bustos²

1998

Journal of Biological Chemistry

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“…L7/L12 ribosomal protein (RP-L7/L12) is among the most investigated components of prokaryotic ribosomes, and it interacts with translation factors during protein biosynthesis in bacteria (24). RP-L7/L12 is present at approximately a 4-fold higher level than other ribosomal proteins, and it increases in proportion to the bacterial growth rate (25).…”

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confidence: 99%

“…Similar proteins are found in the large ribosomal subunits of archaebacteria, eukaryotes, and all eubacteria. Although archaebacterial and eukaryotic proteins are homologous to each other, they show little homology to eubacterial proteins, as assessed by various physical and functional criteria (24). Alignments of the complete RP-L7/L12 amino acid sequences available from 16 different bacterial species show that the C-terminus region is highly conserved; however, one of the monoclonal antibodies (MAbs) cross-reacted only with streptococci in Western blotting (26).…”

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Diagnostic Usefulness of Ribosomal Protein L7/L12 for Pneumococcal Pneumonia in a Mouse Model

et al. 2013

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The capsular antigen detection (CAD) kit is widely used in clinics to detect Streptococcus pneumoniae infection from urine, because it is rapid, convenient, and effective. However, there are several disadvantages, including false-positive results in children colonized with S. pneumoniae and prolonged positive readings even after the bacteria have been cleared. RP-L7/L12 is a component of the 50S ribosome that is abundant in all bacteria and is specific for each bacterial species. We investigated whether RP-L7/L12 could be used to accurately diagnose pneumococcal pneumonia infection in mouse models of pneumonia and colonization generated by infecting CBA/JN or CBA/N mice, respectively, with S. pneumoniae strain 741. RP-L7/L12 detection by enzyme-linked immunosorbent assay accurately assessed active lung infection, as RP-L7/L12 levels decreased simultaneously with the bacterial lung burden after imipenem administration in the pneumonia mouse model. Based on the data, antibodies detecting RP-L7/L12 were applied to rapid immunochromatographic strips (ICS) for urine sample testing. When we compared the ICS test with the CAD kit in the pneumonia model, the results correlated well. Interestingly, however, when the lung bacterial burden became undetectable after antibiotic treatment, the ICS test was correspondingly negative, even though the same samples tested by the CAD kit remained positive. Similarly, while the ICS test exhibited negative results in the nasal colonization model, the CAD kit demonstrated positive results. Bacterial RP-L7/L12 may be a promising target for the development of new methods to diagnose infectious disease. Further studies are warranted to determine whether such a test could be useful in children.

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“…The other important component of the GTPase center is the acidic stalk protein, termed L7/L12 in prokaryotes (11)(12)(13)(14)(15). Four copies of this proteins bind to protein L10 and form a stable complex (16), designated here as L10⅐L7/L12.…”

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confidence: 99%

Translation Elongation by a Hybrid Ribosome in Which Proteins at the GTPase Center of the Escherichia coli Ribosome Are Replaced with Rat Counterparts

Uchiumi¹,

Honma²,

Nomura³

et al. 2002

Journal of Biological Chemistry

105

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Ribosomal L10⅐L7/L12 protein complex and L11 bind to a highly conserved RNA region around position 1070 in domain II of 23 S rRNA and constitute a part of the GTPase-associated center in Escherichia coli ribosomes. We replaced these ribosomal proteins in vitro with the rat counterparts P0⅐P1/P2 complex and RL12, and tested them for ribosomal activities. The core 50 S subunit lacking the proteins on the 1070 RNA domain was prepared under gentle conditions from a mutant deficient in ribosomal protein L11. The rat proteins bound to the core 50 S subunit through their interactions with the 1070 RNA domain. The resultant hybrid ribosome was insensitive to thiostrepton and showed poly(U)-programmed polyphenylalanine synthesis dependent on the actions of both eukaryotic elongation factors 1␣ (eEF-1␣) and 2 (eEF-2) but not of the prokaryotic equivalent factors EF-Tu and EF-G. The results from replacement of either the L10⅐L7/L12 complex or L11 with rat protein showed that the P0⅐P1/P2 complex, and not RL12, was responsible for the specificity of the eukaryotic ribosomes to eukaryotic elongation factors and for the accompanying GTPase activity. The presence of either E. coli L11 or rat RL12 considerably stimulated the polyphenylalanine synthesis by the hybrid ribosome, suggesting that L11/RL12 proteins play an important role in post-GTPase events of translation elongation.The "GTPase center" of the ribosome is a region involved in interaction with GTP-bound translation factors, GTP hydrolysis (1), and post-GTPase events including tRNA movements on the ribosome (2, 3). Translation elongation is markedly stimulated by the interaction of this region with two elongation factors in a GTP-dependent manner. The GTPase center includes two essential RNA regions around positions 1070 and 2660 (Escherichia coli numbering) of the 23 S/28 S rRNA (4), which appear to bind to the elongation factors (5, 6). Despite the highly conserved structure of the 1070 and 2660 RNA regions, ribosomes show a kingdom-dependent accessibility for translation factors, i.e., prokaryotic ribosomes do not engage in translation elongation with the eukaryotic factors instead of the prokaryotic factors (7-9). Furthermore, there are differences in the rate of GTPase turnover between the two systems; in vitro eukaryotic eEF-2 1 /80 S ribosomedependent GTP hydrolysis is 10-fold slower than the prokaryotic EF-G/70 S ribosome system (10). This may reflect, in part, the elaborate regulation of eukaryotic translation.The other important component of the GTPase center is the acidic stalk protein, termed L7/L12 in prokaryotes (11-15). Four copies of this proteins bind to protein L10 and form a stable complex (16), designated here as L10⅐L7/L12. This protein complex and another protein, L11, are assembled on the 1070 RNA domain (17). Flexible property of L7/L12 protein in the ribosome (16, 18 -20) seems to be correlated with the fast turnover of EF-G-dependent GTPase. The eukaryotic counterparts of the prokaryotic L7/L12 and L10 are P1/P2 and P0, respectively (21,22)....

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The L7/L12 ribosomal domain of the ribosome: structural and functional studies

Cited by 59 publications

References 57 publications

Ribosomal P-protein Stalk Function Is Targeted by Sordarin Antifungals

Ribosomal P-protein Stalk Function Is Targeted by Sordarin Antifungals

Diagnostic Usefulness of Ribosomal Protein L7/L12 for Pneumococcal Pneumonia in a Mouse Model

Translation Elongation by a Hybrid Ribosome in Which Proteins at the GTPase Center of the Escherichia coli Ribosome Are Replaced with Rat Counterparts

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