Milón, Pohl

[["dc.bibliographiccitation.firstpage","1"],["dc.bibliographiccitation.lastpage","30"],["dc.bibliographiccitation.volume","430"],["dc.contributor.author","Milon, Pohl"],["dc.contributor.author","Konevega, Andrey L."],["dc.contributor.author","Peske, Frank"],["dc.contributor.author","Fabbretti, Attilio"],["dc.contributor.author","Gualerzi, Claudio O."],["dc.contributor.author","Rodnina, Marina"],["dc.contributor.editor","Ziegler, Christine"],["dc.date.accessioned","2017-09-07T11:49:53Z"],["dc.date.available","2017-09-07T11:49:53Z"],["dc.date.issued","2007"],["dc.description.abstract","Initiation of mRNA translation in prokaryotes requires the small ribosomat subunit (30S), initiator fMet-tRNA(fMet), three initiation factors, IF1, IF2, and IF3, and the large ribosomal subunit (50S). During initiation, the 30S subunit, in a complex with IF3, binds mRNA, IF1, IF2-GTP, and fMet-tRNA(fMet) to form a 30S initiation complex which then recruits the 50S subunit to yield a 70S initiation complex, while the initiation factors are released. Here we describe a transient kinetic approach to study the timing of elemental steps Of 30S initiation complex formation, 50S subunit joining, and the dissociation of the initiation factors from the 70S initiation complex. Labeling of ribosomal subunits, fMet-tRNA(fMet), mRNA, and initiation factors with fluorescent reporter groups allows for the direct observation of the formation or dissociation of complexes by monitoring changes in the fluorescence of single dyes or fluorescence resonance energy transfer (FRET) between two fluorophores. Subunit joining was monitored by light scattering or by FRET between dyes attached to the ribosomat subunits. The kinetics of chemical steps, that is, GTP hydrolysis by IF2 and peptide bond formation following the binding of aminoacyl-tRNA to the 70S initiation complex, were measured by the quench-ftow technique. The methods described here are based on results obtained with initiation components from Escherichia coli but can be adopted for mechanistic studies of initiation in other prokaryotic or eukaryotic systems."],["dc.identifier.doi","10.1016/S0076-6879(07)30001-3"],["dc.identifier.gro","3143569"],["dc.identifier.isi","000250402400001"],["dc.identifier.pmid","17913632"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/1097"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.publisher","Elsevier Academic Press Inc"],["dc.publisher.place","San diego"],["dc.relation.crisseries","Methods in Enzymology"],["dc.relation.isbn","978-0-12-373969-8"],["dc.relation.ispartof","Methods in enzymology"],["dc.relation.ispartofseries","Methods in Enzymology"],["dc.relation.issn","0076-6879"],["dc.title","Transient kinetics, fluorescence, and fret in studies of initiation of translation in bacteria"],["dc.type","book_chapter"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","712"],["dc.bibliographiccitation.issue","6"],["dc.bibliographiccitation.journal","Molecular Cell"],["dc.bibliographiccitation.lastpage","720"],["dc.bibliographiccitation.volume","30"],["dc.contributor.author","Milon, Pohl"],["dc.contributor.author","Konevega, Andrey L."],["dc.contributor.author","Gualerzi, Claudio O."],["dc.contributor.author","Rodnina, Marina V."],["dc.date.accessioned","2017-09-07T11:48:17Z"],["dc.date.available","2017-09-07T11:48:17Z"],["dc.date.issued","2008"],["dc.description.abstract","The translation initiation efficiency of a given mRNA is determined by its translation initiation region (TIR). mRNAs are selected into 30S initiation complexes according to the strengths of the secondary structure of the TIR, the pairing of the Shine-Dalgarno sequence with 16S rRNA, and the interaction between initiator tRNA and the start codon. Here, we show that the conversion of the 30S initiation complex into the translating 70S ribosome constitutes another important mRNA control checkpoint. Kinetic analysis reveals that 50S subunit joining and dissociation of IF3 are strongly influenced by the nature of the codon used for initiation and the structural elements of the TIR. Coupling between the TIR and the rate of 70S initiation complex formation involves IF3- and IF1-induced rearrangements of the 30S subunit, providing a mechanism by which the ribosome senses the TIR and determines the efficiency of translational initiation of a particular mRNA."],["dc.identifier.doi","10.1016/j.molcel.2008.04.014"],["dc.identifier.gro","3143278"],["dc.identifier.isi","000256984300006"],["dc.identifier.pmid","18570874"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/774"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1097-2765"],["dc.title","Kinetic checkpoint at a late step in translation initiation"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]

[["dc.bibliographiccitation.firstpage","312"],["dc.bibliographiccitation.issue","4"],["dc.bibliographiccitation.journal","EMBO reports"],["dc.bibliographiccitation.lastpage","316"],["dc.bibliographiccitation.volume","11"],["dc.contributor.author","Milon, Pohl"],["dc.contributor.author","Carotti, Marcello"],["dc.contributor.author","Konevega, Andrey L."],["dc.contributor.author","Wintermeyer, Wolfgang"],["dc.contributor.author","Rodnina, Marina V."],["dc.contributor.author","Gualerzi, Claudio O."],["dc.date.accessioned","2017-09-07T11:46:06Z"],["dc.date.available","2017-09-07T11:46:06Z"],["dc.date.issued","2010"],["dc.description.abstract","Bacterial translation initiation factor 2 (IF2) is a GTPase that promotes the binding of the initiator fMet-tRNA(fMet) to the 30S ribosomal subunit. It is often assumed that IF2 delivers fMet-tRNA(fMet) to the ribosome in a ternary complex, IF2. GTP. fMet-tRNA(fMet). By using rapid kinetic techniques, we show here that binding of IF2. GTP to the 30S ribosomal subunit precedes and is independent of fMet-tRNA(fMet) binding. The ternary complex formed in solution by IF2. GTP and fMet-tRNA is unstable and dissociates before IF2. GTP and, subsequently, fMet-tRNA(fMet) bind to the 30S subunit. Ribosome-bound IF2 might accelerate the recruitment of fMet-tRNA(fMet) to the 30S initiation complex by providing anchoring interactions or inducing a favourable ribosome conformation. The mechanism of action of IF2 seems to be different from that of tRNA carriers such as EF-Tu, SelB and eukaryotic initiation factor 2 (eIF2), instead resembling that of eIF5B, the eukaryotic subunit association factor."],["dc.identifier.doi","10.1038/embor.2010.12"],["dc.identifier.gro","3142942"],["dc.identifier.isi","000276117100017"],["dc.identifier.pmid","20224578"],["dc.identifier.uri","https://resolver.sub.uni-goettingen.de/purl?gro-2/401"],["dc.language.iso","en"],["dc.notes.intern","WoS Import 2017-03-10"],["dc.notes.status","final"],["dc.notes.submitter","PUB_WoS_Import"],["dc.relation.issn","1469-221X"],["dc.title","The ribosome-bound initiation factor 2 recruits initiator tRNA to the 30S initiation complex"],["dc.type","journal_article"],["dc.type.internalPublication","yes"],["dc.type.peerReviewed","yes"],["dc.type.subtype","original"],["dspace.entity.type","Publication"]]