Antibiotic Inhibition of Protein Synthesis I.PDF

See online version for legend and references.Part II will appear in a new Enhanced SnapShot format in the October 2 issue.1248 Cell 138,September 18,2009 2009 Elsevier Inc.DOI 10.1016/j.cell.2009.08.001SnapShot:Antibiotic Inhibition of Protein Synthesis IDaniel Sohmen,1 Joerg M.Harms,2 Frank Schlnzen,3 and Daniel N.Wilson1,41University of Munich,Germany;2MPSD,University of Hamburg,Germany;3DESY,Hamburg,Germany;4CiPS-M,Munich,Germany ClassNameTargetMechanism Small(30S)Ribosomal SubunitAminoglycosidesApramycin,gentamycin,hygromycin B,kanamycin,neomycin,paromomycin,tobramycinElongation(translocation)Most aminoglycoside antibiotics induce translational misreading by promoting binding of near-cognate tRNAs,but the biological effect is probably due to inhibition of the translocation reaction and promotion of back-translocation.EdeineEdeine AInitiation(fMet-tRNA binding)Edeine prevents binding of the initiator tRNA to the 30S subunit in an mRNA-dependent manner.GE81112GE81112Initiation(fMet-tRNA binding)GE81112 inhibits binding of the initiator tRNA to the 30S subunit in an mRNA-independent manner.KasugamycinKasugamycinInitiation(fMet-tRNA binding)Kasugamycin inhibits translation initiation of canonical mRNAs by binding in the path of the mRNA and preventing stable interaction of the initiator tRNA with the start codon.PactamycinPactamycinInitiation,elongation(translocation)Early data suggest that pactamycin allows 30S but not 70S initiation complex formation,whereas recently pactamycin was shown to inhibit translocation in a tRNA-mRNA-dependent manner.SpectinomycinSpectinomycinElongation(translocation)Spectinomycin binds to the neck of the small subunit and prevents the relative movement of the head and body that is needed for the completion of translocation.StreptomycinStreptomycinElongation(misreading)Streptomycin increases affinity of tRNA for the A site,has a modest inhibitory effect on translocation,promotes back-translocation,and induces high-level translational misreading.Tetracyclines/glycylcyclines Doxycycline,minocycline,tetracycline/tigecyclineElongation(tRNA delivery)Tetracyclines prevent stable binding of the EF-Tu-tRNA-GTP ternary complex to the ribosome and inhibit accommodation of A-tRNAs upon EF-Tu-dependent GTP hydrolysis.ViomycinCapreomycin,viomycinElongation(translocation)Viomycin locks tRNAs in a hybrid-site translocation intermediate state,preventing conversion by EF-G into a posttranslocation state.Large(50S)Ribosomal SubunitBlasticidinsBlasticidin SElongation(peptidyltransferase)Two molecules of Blasticidin S mimic the C74 and C75 of P-tRNA and in doing so inhibit peptide bond formation by preventing tRNA binding at the P site of the peptidyltransferase center(PTC).ChloramphenicolsChloramphenicolElongation(peptidyltransferase)Chloramphenicol binds at the A site of the PTC where it perturbs placement of A site tRNA and thus prevents peptide bond formation.Hygromycin AHygromycin AElongation(peptidyltransferase)Hygromycin A overlaps in its binding site with chloramphenicol and inhibits peptide bond formation by inhibiting the placement of the A site tRNA at the PTC.KetolidesCethromycin(ABT-773),telithromycinElongation(nascent chain egress)Ketolides are derivatives of macrolides where the ketone group replaces the C3 sugar.Ketolides exhibit the same mechanism of action as macrolides by blocking the ribosomal tunnel and inducing peptidyl-tRNA drop-off.LincosamidesClindamycin,lincomycinElongation(peptidyltransferase)Lincosamides span across both A and P sites of the PTC and inhibit binding of tRNA substrates at both these sites.MacrolidesAzithromycin,carbomycin A,clarithromycin,erythromycin,spiramycin,troleandomycin,tylosinElongation(nascent chain egress)Macrolides bind within the ribosomal tunnel and prevent elongation of the nascent polypeptide chain,which leads to peptidyl-tRNA drop-off.Some macrolides,such as carbomycin A and tylosin,also directly inhibit peptide bond formation.OrthosomycinsAvilamycin,evernimicinInitiation(70S-IC formation)Orthosomycins interact with the large subunit and prevent the joining of the 30S-IC with the large subunit to form the 70S-IC in an IF2-dependent manner.OxazolidinonesEperezolid,linezolidElongation(peptidyltransferase)Linezolid binds at the PTC in a position overlapping the aminoacyl moiety of A-tRNA,preventing tRNA accommodation and peptide bond formation.PuromycinPuromycinElongation(peptidyltransferase)Puromycin is a structural analog of the 3 end of a tRNA,which binds at the A site of the PTC and triggers premature release of the nascent polypeptide chain.Ribotoxins-sarcinElongation(translocation),recycling-sarcin cleaves the E.coli 23S rRNA on the 3 side of G2661,abolishing the ribosomes ability to stimulate the GTP hydrolysis activity of translation factors,such as EF-G.SparsomycinSparsomycinElongation(peptidyltransferase)Sparsomycin binds at the PTC where it blocks tRNA binding at the A site,while promoting translocation and stabilization of tRNA at the P site.Streptogramins A/BDalfopristin/quinupristin,virginiamycin M/SElongation(peptidyltransferase)Streptogramins A and B act synergistically to inhibit translation.SA compounds bind at the PTC overlapping A and P sites,whereas SB compounds bind within the tunnel in a similar location to macrolides.ThiopeptidesMicrococcin,nocathiacin,nosiheptide,thiazomycin,thiostreptonElongation(translocation),Recycling Thiopeptide antibiotics inhibit translocation by blocking the stable binding of EF-G to the ribosome.Thiopeptides also inhibit the action of translational GTPases,such IF2 and EF-Tu.Elongation FactorsFusidic acidFusidic acidElongation(translocation)Fusidic acid stabilizes EF-G on the ribosome by allowing binding and GTP hydrolysis,but not Pi release,nor the associated conformational changes in EF-G necessary for dissociation.GE2270A-like thiopeptidesAmythiamicins,GE2270A,thiomuracinsElongation(tRNA delivery)The GE2270A-like thiopeptides,like pulvomycin,prevent ternary complex formation by binding to EF-Tu and blocking interaction of EF-Tu with aminoacyl-tRNAs.GE2270A and pulvomycin have distinct but overlapping binding sites on EF-Tu.Kirromycins and enacyloxinsAurodox,kirromycin,and enacyloxin IIaElongation(tRNA delivery)These drugs trap EF-Tu on the ribosome by allowing GTP hydrolysis but preventing Pi release and the conformational changes in EF-Tu that are necessary for tRNA release and EF-Tu dissociation.Pulvomycin PulvomycinElongation(tRNA delivery)Pulvomycin prevents ternary complex formation by binding to EF-Tu and blocking interaction of EF-Tu with aminoacyl-tRNAs.1248.e1 Cell 138,September 18,2009 2009 Elsevier Inc.DOI 10.1016/j.cell.2009.08.001The translational apparatus is one of the major targets for antibiotics in the bacterial cell.Antibiotics predominantly interact with the functional centers of the ribosome,namely the messenger RNA(mRNA)-transfer RNA(tRNA)decoding region on the 30S subunit,the peptidyltransferase center on the 50S subunit,or the ribosomal exit tunnel through which the nascent polypeptide chain passes during translation.Protein synthesis can be divided into three phases:initiation,elongation,and termination/recycling.1.InitiationTranslation initiation operates through a 30S initiation complex(30S-IC),consisting of the 30S,mRNA,initiator fMet-tRNA,and three initiation factors,IF1,IF2,and IF3.Sub-sequently,the 30S-IC associates with the 50S,which releases the initiation factors(IFs)and leaves the initiator-tRNA at the peptidyl-tRNA-binding site(P site),base-paired to the start codon of the mRNA.The antibiotics edeine,GE81112,and kasugamycin interfere with 30S-IC assembly,whereas formation of a functional 70S-IC is blocked by the orthosomycins and pactamycins.2.ElongationThe elongation phase involves the movement of tRNAs in a cyclic fashion through the three tRNA-binding sites(APE)on the ribosome.The first step in the cycle involves the delivery of the aminoacyl-tRNA(aa-tRNA)to the aa-tRNA-binding site(A site),which is facilitated by the elongation factor EF-TuGTP.Hydrolysis of GTP by EF-Tu leads to its dissociation from the ribosome,allowing aa-tRNA accommodation.Peptide-bond formation then proceeds,transferring the entire polypeptide chain from the P-tRNA to the aa-tRNA in the A site.The ribosome now has a peptidyl-tRNA at the A site and an uncharged tRNA at the P site.This ribosomal state is highly dynamic with the tRNAs oscil-lating between classical(A and P sites)and hybrid states(A/P and P/E sites on 30S/50S).EF-G binds to the ribosome,which stabilizes the tRNAs in hybrid states,hydrolyzes GTP to GDP,and catalyzes the translocation reaction.Translocation shifts the peptidyl-tRNA from the A/P hybrid state to the P site and the deacylated tRNA from the P/E to the exit site(E site).EF-GGDP dissociates leaving the A site vacant for the next incoming aa-tRNA.The majority of antibiotics targeting translation inhibit a step associated with the elongation phase.Most antibiotics binding to the 30S perturb either tRNA binding,decoding,or translocation,whereas antibiotics targeting the 50S inhibit peptide-bond formation,extension of the polypeptide chain,or stable binding of translation factors.In contrast,some antibiotics interact directly with elongation factors and prevent them from dissociating from the ribosome.3.Termination/RecyclingArrival of an mRNA stop codon in the A site of the ribosome signals the termination of protein synthesis.Release factor 1(RF1)or RF2 binds to the ribosome and hydrolyzes the peptidyl-tRNA bond,releasing the translated polypeptide chain from the ribosome.RF1/2 is recycled from the ribosome by RF3 in a GTP-dependent fashion.The ribosome is then split into subunits by the concerted action of EF-G and ribosome recycling factor(RRF),thus recycling the components for the next round of translation.There are no antibiotics that specifically inhibit termination and ribosome recycling;however antibiotics that target EF-G,such as-sarcin,fusidic acid,and thiopeptides,also inhibit ribo-some recycling.Abbreviations30S,small ribosomal subunit;50S,large ribosomal subunit;aa-tRNA,aminoacyl-tRNA;A site,aa-tRNA-binding site;E site,exit site for tRNA;EF,elongation factor;IF,initiation factor;mRNA,messenger RNA;P-tRNA,peptidyl-tRNA;P site,P-tRNA-binding site;PTC,peptidyltransferase center;RRF,ribosome recycling factor;RF,release factor;tRNA,transfer RNA.ACkNowLEdgMENTSD.N.W.is funded by the Deutsche Forschungsgemeinschaft(WI3285/1-1)and HFSP(RGY0088/2008).REFERENCESMoore,P.,and Steitz,T.(2003).The structural basis of large ribosomal subunit function.Annu.Rev.Biochem.72,813850.Parmeggiani,A.,and Nissen,P.(2006).Elongation factor Tu-targeted antibiotics:four different structures,two mechanisms of action.FEBS Lett.580,45764581.Poehlsgaard,J.,and Douthwaite,S.(2005).The bacterial ribosome as a target for antibiotics.Nat.Rev.Microbiol.3,870881.Polacek,N.,and 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Munich,Germany;2MPSD,University of Hamburg,Germany;3DESY,Hamburg,Germany;4CiPS-M,Munich,Germany