Wintermeyer, Wolfgang

[["dc.bibliographiccitation.firstpage","301"],["dc.bibliographiccitation.lastpage","318"],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.contributor.author","Savelsbergh, Andreas"],["dc.contributor.author","Pape, Tillmann"],["dc.contributor.author","Mohr, Dagmar"],["dc.contributor.editor","Garrett, Roger A."],["dc.date.accessioned","2017-09-07T11:54:06Z"],["dc.date.available","2017-09-07T11:54:06Z"],["dc.date.issued","2014"],["dc.description.abstract","This chapter concentrates on pre-steady-state kinetic work, and provides evidence on how much the interpretation of kinetic results in molecular-mechanistic terms owes to structural information obtained from crystallography and cryo-electron microscopy. Our studies of elongation factor (EF-Tu) function address two main issues: (i) the elucidation of the reaction pathway to identify intermediate steps of A-site binding and (ii) the quantitative evaluation of the pathway in order to understand specificity. Based on measured rates of GTP hydrolysis and peptide bond formation, Thompson and colleagues proposed that the rate of GTP hydrolysis by EF-Tu is independent of the tRNA, thereby providing an internal kinetic standard for translational accuracy. Binding of thiostrepton to the 1070 region of 23S rRNA interferes with translocation, as it strongly inhibits Pi release, translocation, and subsequent turnover of EF-G; in contrast, EF-G binding and GTP hydrolysis are not affected. The GTPase activities of EF-Tu and EF-G intrinsically are very low and are strongly enhanced on the ribosome. Recently, the ability of isolated L12 protein to stimulate GTP hydrolysis by either EF-Tu or EF-G, has been studied. In the fusidic acid-stabilized complex of EF-G with the ribosome, the α- sarcin stem is close to position 196 in the G domain of EF-G, which lies just above the GTP binding site, while the α-sarcin loop region is in the vicinity of position 650 in domain 5 of the factor."],["dc.identifier.doi","10.1128/9781555818142.ch25"],["dc.identifier.gro","3145105"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2805"],["dc.language.iso","en"],["dc.notes.intern","Crossref Import"],["dc.notes.status","final"],["dc.publisher","American Society for Microbiology Press"],["dc.publisher.place","Washington, DC"],["dc.relation.doi","10.1128/9781555818142"],["dc.relation.isbn","1-55581-184-1"],["dc.relation.isbn","978-1-55581-184-6"],["dc.relation.ispartof","The Ribosome: Structure, Function, Antibiotics, and Cellular Interactions"],["dc.title","Mechanisms of Partial Reactions of the Elongation Cycle Catalyzed by Elongation Factors Tu and G"],["dc.type","book_chapter"],["dc.type.internalPublication","no"],["dc.type.peerReviewed","no"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","1879"],["dc.bibliographiccitation.issue","12"],["dc.bibliographiccitation.journal","RNA"],["dc.bibliographiccitation.lastpage","1885"],["dc.bibliographiccitation.volume","7"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:46:01Z"],["dc.date.available","2017-09-07T11:46:01Z"],["dc.date.issued","2001"],["dc.description.abstract","During the translocation step of the elongation cycle, two tRNAs together with the mRNA move synchronously and rapidly on the ribosome. The movement is catalyzed by the binding of elongation factor G (EF-G) and driven by GTP hydrolysis. Here we study structural changes of the ribosome related to! EF-G binding and translocation by monitoring the accessibility of ribosomal RNA (rRNA) for chemical modification by dimethyl sulfate or cleavage by hydroxyl radicals generated by Fe(II)-EDTA. In the state of the ribosome that is formed upon binding of EF-G but before the movement of the tRNAs takes place, residues 1054, 1196, and 1201 in helix 34 in 16S rRNA are strongly protected. The protections depend on EF-G binding, but do not require GTP hydrolysis, and are lost upon translocation. Mutants of EF-G, which are active in ribosome binding and GTP hydrolysis but impaired in translocation, do not bring about the protections. According to cryo-electron microscopy (Stark et al., Cell, 2000, 100:301-309), there is no contact of EF-G with the protected residues of helix 34 in the pretranslocation state,, suggesting that the observed protections are due to an induced conformational change. Thus, the present results indicate that EF-G binding to them pretranslocation ribosome induces a structural change of the: head of the 30S subunit that is essential for subsequent tRNA-mRNA movement in translocation."],["dc.identifier.gro","3144242"],["dc.identifier.isi","000172966300018"],["dc.identifier.pmid","11780642"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1844"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Cambridge Univ Press"],["dc.relation.issn","1355-8382"],["dc.title","Elongation factor G-induced structural change in helix 34 of 16S rRNA related to translocation on the ribosome"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","377"],["dc.bibliographiccitation.issue","5-6"],["dc.bibliographiccitation.journal","Biological Chemistry"],["dc.bibliographiccitation.lastpage","387"],["dc.bibliographiccitation.volume","381"],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Stark, Holger"],["dc.contributor.author","Savelsbergh, Andreas"],["dc.contributor.author","Wieden, Hans-Joachim"],["dc.contributor.author","Mohr, Dagmar"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Peske, Frank"],["dc.contributor.author","Daviter, T"],["dc.contributor.author","Gualerzi, Claudio O."],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:47:23Z"],["dc.date.available","2017-09-07T11:47:23Z"],["dc.date.issued","2000"],["dc.description.abstract","The elongation factors (EF) Tu and G and initiation factor 2 (IF2) from bacteria are multidomain GTPases with essential functions in the elongation and initiation phases of translation. They bind to the same site on the ribosome where their low intrinsic GTPase activities are strongly stimulated. The factors differ fundamentally from each other, and from the majority of GTPases, in the mechanisms of GTPase control, the timing of P-i release, and the functional role of GTP hydrolysis. EF-Tu GTP forms a ternary complex with aminoacyl-tRNA, which binds to the ribosome, Only when a matching codon is recognized, the GTPase of EF-Tu is stimulated, rapid GTP hydrolysis and P-i release take place, EF-Tu rearranges to the GDP form, and aminoacyl-tRNA is released into the peptidyltransferase center. In contrast, EF-G hydrolyzes GTP immediately upon binding to the ribosome, stimulated by ribosomal protein L7/12. Subsequent translocation is driven by the slow dissociation of P-i, suggesting a mechano-chemical function of EF-G, Accordingly, different conformations of EF-G on the ribosome are revealed by cryo-electron microscopy. GTP hydrolysis by IF2 is triggered upon formation of the 70S initiation complex, and the dissociation of P-i and/or IF2 follows a rearrangement of the ribosome into the elongation-competent state."],["dc.identifier.doi","10.1515/BC.2000.050"],["dc.identifier.gro","3144395"],["dc.identifier.isi","000088435900004"],["dc.identifier.pmid","10937868"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2015"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Walter De Gruyter & Co"],["dc.relation.issn","1431-6730"],["dc.title","GTPase mechanisms and functions of translation factors on the ribosome"],["dc.type","review"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","951"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","Journal of Molecular Biology"],["dc.bibliographiccitation.lastpage","961"],["dc.bibliographiccitation.volume","300"],["dc.contributor.author","Savelsbergh, Andreas"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:46:48Z"],["dc.date.available","2017-09-07T11:46:48Z"],["dc.date.issued","2000"],["dc.description.abstract","Elongation factor G (EF-G) is a large, five domain GTPase that catalyses the translocation of the tRNAs on the bacterial ribosome at the expense of GTP. in the crystal structure of GDP-bound EF-G, domain 1 (G domain) makes direct contacts with domains 2 and 5, whereas domain 4 protrudes from the body of the molecule. Here, we show that the presence of both domains 4 and 5 is essential for tRNA translocation and for the turnover of the factor on the ribosome, but not for rapid single-round GTP hydrolysis by EF-G. Replacement of a highly conserved histidine residue at the tip of domain 4, His583, with lysine or arginine decreases the rate of tRNA translocation at least 100-fold, whereas the binding of the factor to the ribosome, GTP hydrolysis and P-i release are not affected by the mutations. Various small deletions in the tip region of domain 4 decrease the translocation activity of EF-G even further, but do not block the turnover of the factor. Unlike native EF-G, the mutants of EF-G lacking domains 4/5 do not interact with the alpha-sarcin stem-loop of 23 S rRNA. These mutants are not released from the ribosome after GTP hydrolysis or translocation, indicating that the contact with, or a conformational change of, the alpha-sarcin stem-loop is required for EF-G release from the ribosome. (C) 2000 Academic Press."],["dc.identifier.doi","10.1006/jmbi.2000.3886"],["dc.identifier.gro","3144370"],["dc.identifier.isi","000088508500021"],["dc.identifier.pmid","10891280"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1986"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Academic Press Ltd"],["dc.relation.issn","0022-2836"],["dc.title","Role of domains 4 and 5 in elongation factor G functions on the ribosome"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","939"],["dc.bibliographiccitation.issue","7"],["dc.bibliographiccitation.journal","RNA"],["dc.bibliographiccitation.lastpage","946"],["dc.bibliographiccitation.volume","5"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Endermann, R."],["dc.contributor.author","Kroll, HP"],["dc.contributor.author","Pleiss, U."],["dc.contributor.author","Wildhagen, Henning"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:47:32Z"],["dc.date.available","2017-09-07T11:47:32Z"],["dc.date.issued","1999"],["dc.description.abstract","Oxazolidinones are antibacterial agents that act primarily against gram-positive bacteria by inhibiting protein synthesis. The binding of oxazolidinones to 70S ribosomes from Escherichia coli was studied by both UV-induced cross-linking using an azido derivative of oxazolidinone and chemical footprinting using dimethyl sulphate. Oxazolidinone binding sites were found on both 30S and 50S subunits, rRNA being the only target. On 16S rRNA, an oxazolidinone footprint was found at A864 in the central domain. 23S rRNA residues involved in oxazolidinone binding were U2113, A2114, U2118, A2119, and C2153, all in domain V. This region is close to the binding site of protein L1 and of the 3' end of tRNA in the E site. The mechanism of action of oxazolidinones in vitro was examined in a purified translation system from E. coli using natural mRNA. The rate of elongation reaction of translation was decreased, most probably because of an inhibition of tRNA translocation, and the length of nascent peptide chains was strongly reduced. Both binding sites and mode of action of oxazolidinones are unique among the antibiotics known to act on the ribosome."],["dc.identifier.doi","10.1017/S1355838299990210"],["dc.identifier.gro","3144465"],["dc.identifier.isi","000081268600010"],["dc.identifier.pmid","10411137"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2092"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Cambridge Univ Press"],["dc.relation.issn","1355-8382"],["dc.title","Ribosomal RNA is the target for oxazolidinones, a novel class of translational inhibitors"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","9586"],["dc.bibliographiccitation.issue","17"],["dc.bibliographiccitation.journal","Proceedings of the National Academy of Sciences"],["dc.bibliographiccitation.lastpage","9590"],["dc.bibliographiccitation.volume","96"],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Savelsbergh, Andreas"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Katunin, Vladimir I."],["dc.contributor.author","Semenkov, Yuri P."],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:47:28Z"],["dc.date.available","2017-09-07T11:47:28Z"],["dc.date.issued","1999"],["dc.description.abstract","The region around position 1067 in domain II of 23S rRNA frequently is referred to as the GTPase renter of the ribosome, The notion is based on the observation that the binding of the antibiotic thiostrepton to this region inhibited GTP hydrolysis by elongation factor G (EF-G) on the ribosome at the conditions of multiple turnover, In the present work, we have reanalyzed the mechanism of action of thiostrepton, Results obtained by biochemical and fast kinetic techniques show that thiostrepton binding to the ribosome does not interfere with factor binding or with single-round GTP hydrolysis, Rather, the antibiotic inhibits the function of EF-G in subsequent steps, including release of inorganic phosphate from EF-G after GTP hydrolysis, tRNA translocation, and the dissociation of the factor from the ribosome, thereby inhibiting the turnover reaction. Structurally. thiostrepton interferes with EF-G footprints in the alpha-sarcin stem loop (A2660, A2662) located in domain VI of 23S rRNA. The results indicate that thiostrepton inhibits a structural transition of the 1067 region of 23S rRNA that is important for functions of EF-G after GTP hydrolysis."],["dc.identifier.doi","10.1073/pnas.96.17.9586"],["dc.identifier.gro","3144448"],["dc.identifier.isi","000082098500027"],["dc.identifier.pmid","10449736"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/2073"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Natl Acad Sciences"],["dc.relation.issn","0027-8424"],["dc.title","Thiostrepton inhibits the turnover but not the GTPase of elongation factor G on the ribosome"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","293"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","RNA"],["dc.bibliographiccitation.lastpage","301"],["dc.bibliographiccitation.volume","7"],["dc.contributor.author","Jagath, J. R."],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Leeuw, E. de"],["dc.contributor.author","Warnecke, J. M."],["dc.contributor.author","Lentzen, G."],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Luirink, J"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:46:41Z"],["dc.date.available","2017-09-07T11:46:41Z"],["dc.date.issued","2001"],["dc.description.abstract","Binding of Escherichia coli signal recognition particle (SRP) to its receptor, FtsY, requires the presence of 4.5S RNA, although FtsY alone does not interact with 4.5S RNA. In this study, we report that the exchange of the GGAA tetraloop sequence in domain IV of 4.5S RNA for UUCG abolishes SRP-FtsY interaction, as determined by gel retardation and membrane targeting experiments, whereas replacements with other GNRA-type tetraloops have no effect. A number of other base exchanges in the tetraloop sequence have minor or intermediate inhibitory effects. Base pair disruptions in the stem adjacent to the tetraloop or replacement of the closing C-G base pair with G-C partially restored function of the otherwise inactive UUCG mutant. Chemical probing by hydroxyl radical cleavage of 4.5S RNA variants show that replacing GGAA with UUCG in the tetraloop sequence leads to structural changes both within the tetraloop and in the adjacent stem; the latter change is reversed upon reverting the C-G closing base pair to G-C, These results show that the SRP-FtsY interaction is strongly influenced by the structure of the tetraloop region of SRP RNA, in particular the tetraloop stem, and suggest that both SRP RNA and Ffh undergo mutual structural adaptation to form SRP that is functional in the interaction with the receptor, FtsY."],["dc.identifier.doi","10.1017/S1355838201002205"],["dc.identifier.gro","3144309"],["dc.identifier.isi","000166884500014"],["dc.identifier.pmid","11233986"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1919"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Cambridge Univ Press"],["dc.relation.issn","1355-8382"],["dc.title","Important role of the tetraloop region of 4.5S RNA in SRP binding to its receptor FtsY"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","501"],["dc.bibliographiccitation.issue","2"],["dc.bibliographiccitation.journal","Molecular Cell"],["dc.bibliographiccitation.lastpage","505"],["dc.bibliographiccitation.volume","6"],["dc.contributor.author","Peske, Frank"],["dc.contributor.author","Matassova, Natalia B."],["dc.contributor.author","Savelsbergh, Andreas"],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.date.accessioned","2017-09-07T11:46:48Z"],["dc.date.available","2017-09-07T11:46:48Z"],["dc.date.issued","2000"],["dc.description.abstract","Elongation factor G (EF-G) from Escherichia coli is a large, five-domain GTPase that promotes tRNA translocation on the ribosome. Full activity requires GTP hydrolysis, suggesting that a conformational change of the factor is important for function. To restrict the intramolecular mobility, two cysteine residues were engineered into domains 1 and 5 of EF-G that spontaneously formed a disulfide cross-link. Cross-linked EF-G retained GTPase activity on the ribosome, whereas it was inactive in translocation as well as in turnover. Both activities were restored when the cross-link was reversed by reduction. These results strongly argue against a GTPase switch-type model of EF-G function and demonstrate that conformational mobility is an absolute requirement for EF-G function on the ribosome."],["dc.identifier.doi","10.1016/S1097-2765(00)00049-6"],["dc.identifier.gro","3144367"],["dc.identifier.isi","000089166100027"],["dc.identifier.pmid","10983996"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1983"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Cell Press"],["dc.relation.issn","1097-2765"],["dc.title","Conformationally restricted elongation factor G retains GTPase activity but is inactive in translocation on the ribosome"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]