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Cited by 451Antibiotic Resistance in BacteriaRNA and protein synthesis mechanismsBacterial Genetics and Biotechnology

Ribosomal Protection Proteins and Their Mechanism ofTetracyclineResistance

Open access
Connell, Sean R., Tracz, Dobryan M., Nierhaus, Knud H., Taylor, Diane E.·2003·Antimicrobial Agents and Chemotherapy
DOI 10.1128/aac.47.12.3675-3681.2003PMC PMC296194
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Abstract

Ribosomal protection represents an important tactic for promoting tetracycline resistance in both gram-positive and -negative species. Tet(O) and Tet(M) are the best studied of these determinants and were originally isolated from Campylobacter jejuni and Streptococcus spp., respectively, although both are widely distributed (10). These are the only two ribosomal protection proteins (RPPs) that have been studied in detail, and therefore, they have been dealt with extensively in this review. It is assumed, however, that the other members of this class of RPPs [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through similar mechanisms. The distribution of these determinants in the eubacteria has been extensively reviewed by Chopra and Roberts (10) and more recent information can also be found at http://faculty.washington.edu/marilynr/. Although this review focuses primarily on RPPs, it should be noted that a great variety of tetracycline resistance mechanisms exist (for a review, see reference 10). These determinants include (i) the efflux-based mechanisms found in gram-positive and gram-negative bacteria (10), (ii) the enzymatic degradation of tetracyclines found in Bacteroides (46), (iii) the rRNA mutations found in Propionibacterium acnes and Helicobacter pylori (19, 40, 55), and (iv) a host of undetermined mechanisms which bear little resemblance to the well-documented determinants mentioned above (10). In this review, we will survey recent advances in the study of the ribosome, tetracycline, and the RPPs that further the understanding of RPP activity. Earlier work dealing with Tet(M) and Tet(O) as well as the other RPPs has been reviewed previously (51, 52).

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Written from the paper’s full text.

TL;DR

Ribosomal protection proteins (RPPs) such as Tet(O) and Tet(M) confer tetracycline resistance by binding the ribosome in a GTP-dependent manner and releasing tetracycline from the primary 30S binding site, restoring A-site tRNA accommodation. The review integrates biochemical and structural data into an allosteric model in which Tet(O) contacts h34 and perturbs the decoding center.

Key findings

  • Tetracycline inhibits elongation by blocking aminoacyl-tRNA accommodation into the ribosomal A site; the high-affinity 30S binding site has a \(K_d\) of 1 to 20 µM, while crystal structures show tetracycline bound to either two or six sites on the 30S subunit. The primary/Tet-1 site near h34 and h31 is the likely inhibitory site.
  • Tet(O) and Tet(M) are soluble cytoplasmic RPPs of ~72 kDa with 75% sequence similarity; they dislodge tetracycline from the ribosome and increase the apparent \(K_d\) of tetracycline binding from 5 to 30 µM, and Tet(M) protects A-site tRNA binding from tetracycline inhibition.
  • RPPs bind the elongation factor site on the ribosome. A 16-Å cryo-EM reconstruction shows Tet(O) at the subunit interface, contacting mostly rRNA; its domain IV contacts h18/34 of 16S rRNA, unlike EF-G domain IV, which contacts H69 of 23S rRNA. DMS probing shows Tet(O) protects C1214 and enhances A1408.
  • Proposed mechanism: Tet(O) binds a tetracycline-blocked ribosome with an open A site, triggers allosteric release of tetracycline via disturbance of h34, hydrolyzes GTP, and dissociates, returning the ribosome to the elongation cycle.

Methods

  • Minireview synthesizing published biochemical, chemical probing, cryo-EM, and crystal structure studies; no new experimental data are reported.
  • Focuses on Tet(O) and Tet(M), the only two RPPs studied in detail, and assumes other RPPs [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through similar mechanisms.
  • Main analysis integrates ribosome-tetracycline structures and RPP binding data into a model of tetracycline action and RPP-mediated tetracycline release.

Limitations

  • Stated: only Tet(O) and Tet(M) have been studied in detail; other RPPs are assumed to function similarly.
  • Stated: it remains unanswered whether RPPs actively prevent tetracycline rebinding after release; also, there is a discrepancy about GTP hydrolysis, as a nonhydrolyzable GTP analogue was active with Tet(O) but only partially active with Tet(M).
  • Evident from the text: several mechanistic steps, including a tetracycline-induced conformational change and the role of h44 rearrangement, are proposed but not conclusively established.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1128/aac.47.12.3675-3681.2003

Cites

60 of 60 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 103 cited sources checked · 0 supported · 0 flagged

  • Premisecitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    The face of tetracycline that interacts with rRNA in the primary (and Tet-1) site is also the face where modifications result in a loss of biological activity.

    “(iii) The crystal structures show that the face of tetracycline that interacts with the rRNA in the primary (and Tet-1) site is also the face where modifications result in a loss of biological activity (4, 39).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [4] 4. Brodersen, D. E., W. M. Clemons, A. P. Carter, R. J. Morgan-Warren, B. T. Wimberly, and V. Ramakrishnan. 2000. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell 103: 1143-1154. source

      Not checked in the free run.

    • cites [39] 39. Rogalski, W. 1985. Chemical modification of the tetracyclines, p. 179-316. In J. H. Boothe and J. J. Hlavka (ed.), The tetracyclines (handbook of experimental pharmacology), vol. 78. Springer-Verlag, Berlin, Germany. source

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  • Interpretationreasoning
    § RIBOSOMAL PROTECTION PROTEINS

    RPPs may be evolutionarily derived from elongation factors, such that they lost their original function and have been adapted to function in tetracycline resistance.

    “Nevertheless, the RPPs may be evolutionarily derived from the elongation factors, such that they lost their original function and have been adapted to function in tetracycline resistance.”
    Quote in the paperNot a substantial claim
  • Premisecitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    Two independent 30S subunit-tetracycline crystal structures show tetracycline bound to either two or six sites on the 30S ribosomal subunit.

    “This is significant, as two independent 30S subunit-tetracycline crystal structures show tetracycline bound to either two (4) or six sites (37) on the 30S ribosomal subunit (Fig. 2).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [4] 4. Brodersen, D. E., W. M. Clemons, A. P. Carter, R. J. Morgan-Warren, B. T. Wimberly, and V. Ramakrishnan. 2000. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell 103: 1143-1154. source

      Not checked in the free run.

    • cites [37] 37. Pioletti, M., F. Schlunzen, J. Harms, R. Zarivach, M. Gluhmann, H. Avila, A. Bashan, H. Bartels, T. Auerbach, C. Jacobi, T. Hartsch, A. Yonath, and F. Franceschi. 2001. Crystal structures of complexes of the small ribosomal subunit with tetracycline, edeine and IF3. EMBO J. 20: 1829-1839. source

      Not checked in the free run.

  • Interpretationcitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    The secondary and Tet-5 tetracycline-binding sites are likely candidates for the inhibitory binding site.

    “The secondary and Tet-5 tetracycline-binding sites (Fig. 2) are also likely candidates for the inhibitory binding site (4, 37).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [4] 4. Brodersen, D. E., W. M. Clemons, A. P. Carter, R. J. Morgan-Warren, B. T. Wimberly, and V. Ramakrishnan. 2000. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell 103: 1143-1154. source

      Not checked in the free run.

    • cites [37] 37. Pioletti, M., F. Schlunzen, J. Harms, R. Zarivach, M. Gluhmann, H. Avila, A. Bashan, H. Bartels, T. Auerbach, C. Jacobi, T. Hartsch, A. Yonath, and F. Franceschi. 2001. Crystal structures of complexes of the small ribosomal subunit with tetracycline, edeine and IF3. EMBO J. 20: 1829-1839. source

      Not checked in the free run.

  • Premisecitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    In the primary (and Tet-1) site, several tetracycline-RNA interactions are mediated through a magnesium ion known to be important for tetracycline binding.

    “(ii) In the primary (and Tet-1) site, several tetracycline-RNA interactions are mediated through a magnesium ion which is known to be important for tetracycline binding (4, 57).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [4] 4. Brodersen, D. E., W. M. Clemons, A. P. Carter, R. J. Morgan-Warren, B. T. Wimberly, and V. Ramakrishnan. 2000. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell 103: 1143-1154. source

      Not checked in the free run.

    • cites [57] 57. White, J. P., and C. R. Cantor. 1971. Role of magnesium in the binding of tetracycline to Escherichia coli ribosomes. J. Mol. Biol. 58: 397-400. source

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  • May combine two claims. 18
  • Classified as probably not a substantial claim. 10

  • May combine two claims.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    Many RPP determinants are located on mobile genetic elements, which may have facilitated their spread throughout the eubacteria via lateral gene transfer.

    “Accordingly, many of the RPP determinants are located on mobile genetic elements which may have facilitated their spread throughout the eubacteria via lateral gene transfer events (reviewed in reference 10).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [10] 10. Chopra, I., and M. Roberts. 2001. Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. Microbiol. Mol. Biol. Rev. 65: 232-260. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    RPPs bind and hydrolyze GTP in a ribosome-dependent manner, and maintenance of this activity is important for in vivo activity.

    “Accordingly, the RPPs bind and hydrolyze GTP in a ribosome-dependent manner (5, 6, 53), and maintenance of this activity is important for in vivo activity (11, 23).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [5] 5. Burdett, V. 1991. Purification and characterization of Tet(M), a protein that renders ribosomes resistant to tetracycline. J. Biol. Chem. 266: 2872-2877. source

      Not checked in the free run.

    • cites [6] 6. Burdett, V. 1996. Tet(M)-promoted release of tetracycline from ribosomes is GTP dependent. J. Bacteriol. 178: 3246-3251. source

      Not checked in the free run.

    • cites [53] 53. Taylor, D. E., L. J. Jerome, J. Grewal, and N. Chang. 1995. Tet(O), a protein that mediates ribosomal protection to tetracycline, binds, and hydrolyses GTP. Can. J. Microbiol. 41: 965-970. source

      Not checked in the free run.

    • The authors state this as an assumption: The other members of the RPP class [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through mechanisms similar to those of Tet(O) and Tet(M).

  • May combine two claims.

    Premisecitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    Typical tetracyclines bind to the ribosome and inhibit the elongation phase of protein synthesis.

    “Alternatively, the typical tetracyclines, which are the subject of RPP-mediated resistance, bind to the ribosome and inhibit the elongation phase of protein synthesis (8, 13).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [8] 8. Chopra, I. 1985. Mode of action of the tetracyclines and the nature of bacterial resistance to them, p. 317-392. In J. H. Boothe and J. J. Hlavka (ed.), The tetracyclines (handbook of experimental pharmacology), vol. 78. Springer-Verlag, Berlin, Germany. source

      Not checked in the free run.

    • cites [13] 13. Cundliffe, E. 1967. Antibiotics and polyribosomes. Chlortetracycline and polyribosomes of Bacillus megaterium . Mol. Pharmacol. 3: 401-411.

      Not checked in the free run.

  • May combine two claims.

    Premisecitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    Tet(O) chases tetracyclines from the primary binding site but not the secondary binding site.

    “(vi) Tet(O), an RPP which confers resistance to tetracycline, chases tetracyclines from the primary binding site but not the secondary binding site (12).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [12] 12. Connell, S. R., C. A. Trieber, U. Stelzl, E. Einfeldt, D. E. Taylor, and K. H. Nierhaus. 2002. The tetracycline resistance protein, Tet(O), perturbs the conformation of the ribosomal decoding center. Mol. Microbiol. 45: 14631472. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    The primary (and Tet-1) site is the most highly occupied site in both structures showing tetracycline bound to the 30S subunit, consistent with the high-affinity site being the inhibitory site.

    “(i) the primary (and Tet-1) site is the most highly occupied site in both structures showing tetracycline bound to the 30S subunit (4, 37) consistent with the idea that the high-affinity site is the inhibitory site (17, 47, 56).”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [4] 4. Brodersen, D. E., W. M. Clemons, A. P. Carter, R. J. Morgan-Warren, B. T. Wimberly, and V. Ramakrishnan. 2000. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell 103: 1143-1154. source

      Not checked in the free run.

    • cites [37] 37. Pioletti, M., F. Schlunzen, J. Harms, R. Zarivach, M. Gluhmann, H. Avila, A. Bashan, H. Bartels, T. Auerbach, C. Jacobi, T. Hartsch, A. Yonath, and F. Franceschi. 2001. Crystal structures of complexes of the small ribosomal subunit with tetracycline, edeine and IF3. EMBO J. 20: 1829-1839. source

      Not checked in the free run.

    • cites [17] 17. Epe, B., and P. Woolley. 1984. The binding of 6-demethylchlortetracycline to 70S, 50S and 30S ribosomal particles: a quantitative study by fluorescence anisotropy. EMBO J. 3: 121-126. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    RPPs such as Tet(O) and Tet(M) are soluble cytoplasmic proteins of approximately 72 kDa that mediate tetracycline resistance.

    “RPPs, such as the well-studied Tet(O) and Tet(M) (75% sequence similarity), are soluble cytoplasmic proteins ( ∼ 72 kDa) which mediate tetracycline resistance (51).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [51] 51. Taylor, D. E., and A. Chau. 1996. Tetracycline resistance mediated by ribosomal protection. Antimicrob. Agents Chemother. 40: 1-5. source

      Not checked in the free run.

    • The authors state this as an assumption: The other members of the RPP class [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through mechanisms similar to those of Tet(O) and Tet(M).

  • May combine two claims.

    Premisecitation
    § INHIBITORY ACTIONS OF TETRACYCLINES

    Typical tetracyclines inhibit accommodation of aminoacyl-tRNA into the ribosomal A site and prevent addition of new amino acids to the growing polypeptide.

    “More precisely, they inhibit accommodation of aminoacyl-tRNA (aa-tRNA) into the ribosomal A site (Fig. 1, reactions a and b) and, therefore, prevent the addition of new amino acids to the growing polypeptide (22, 25, 29, 48).”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [22] 22. Gordon, J. 1969. Hydrolysis of guanosine 5 ′ -triphosphate associated with binding of aminoacyl transfer ribonucleic acid to ribosomes. J. Biol. Chem. 244: 5680-5686. source

      Not checked in the free run.

    • cites [25] 25. Hierowski, M. 1965. Inhibition of protein synthesis by chlorotetracycline in the Escherichia coli in vitro system. Proc. Natl. Acad. Sci. USA 53: 594-599. source

      Not checked in the free run.

    • cites [29] 29. Lucas-Lenard, J., and A. L. Haenni. 1967. Requirement of guanosine 5 ′ -triphosphate for ribosomal binding of aminoacyl-sRNA. Biochemistry 59: 554-560.

      Not checked in the free run.

    • cites [48] 48. Suarez, G., and D. Nathans. 1965. Inhibition of aminoacyl tRNA binding to ribosomes by tetracycline. Biochem. Biophys. Res. Commun. 18: 743-750. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    Tet(O) was first cloned from a transferable plasmid pUA466 found in the food-borne pathogen C. jejuni.

    “Tet(O) was first cloned from a transferable plasmid pUA466 found in the food-borne pathogen C. jejuni (50).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [50] 50. Taylor, D. E. 1986. Plasmid-mediated tetracycline resistance in Campylobacter jejuni : expression in Escherichia coli and identification of homology with streptococcal class M determinant. J. Bacteriol. 165: 1037-1039. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    Tet(O) and Tet(M) have 75% sequence similarity.

    “RPPs, such as the well-studied Tet(O) and Tet(M) (75% sequence similarity), are soluble cytoplasmic proteins ( ∼ 72 kDa) which mediate tetracycline resistance (51).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [51] 51. Taylor, D. E., and A. Chau. 1996. Tetracycline resistance mediated by ribosomal protection. Antimicrob. Agents Chemother. 40: 1-5. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    RPPs display sequence similarity to EF-G and EF-Tu and are grouped into the translation factor superfamily of GTPases.

    “The RPPs display sequence similarity to the ribosomal elongation factors, EF-G and EF-Tu (41), and are grouped into the translation factor superfamily of GTPases (27).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [41] 41. Sanchez-Pescador, R., J. T. Brown, M. Roberts, and M. S. Urdea. 1988. Homology of the TetM with translational elongation factors: implications for potential modes of tetM -conferred tetracycline resistance. Nucleic Acids Res. 16: 1218. source

      Not checked in the free run.

    • cites [27] 27. Leipe, D. D., Y. I. Wolf, E. V. Koonin, and L. Aravind. 2002. Classification and evolution of P-loop GTPases and related ATPases. J. Mol. Biol. 317: 41-72. source

      Not checked in the free run.

    • The authors state this as an assumption: The other members of the RPP class [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through mechanisms similar to those of Tet(O) and Tet(M).

  • May combine two claims.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    These changes could be mediated by S12, which is in close proximity to the top of h44 and also appears to interact with Tet(O).

    “These changes could be mediated by S12, which is in close proximity to the top of h44 (43, 58) and also appears to interact with Tet(O) (45).”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [43] 43. Schluenzen, F., A. Tocilj, R. Zarivach, J. Harms, M. Gluehmann, D. Janell, A. Bashan, H. Bartels, I. Agmon, F. Franceschi, and A. Yonath. 2000. Structure of functionally activated small ribosomal subunit at 3.3 angstroms resolution. Cell 102: 615-623. source

      Not checked in the free run.

    • cites [58] 58. Wimberly, B. T., D. E. Brodersen, W. M. Clemons, Jr., R. J. Morgan-Warren, A. P. Carter, C. Vonrhein, T. Hartsch, and V. Ramakrishnan. 2000. Structure of the 30S ribosomal subunit. Nature 407: 327-339. source

      Not checked in the free run.

    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    Tet(O) triggers the release of tetracycline through an allosteric mechanism.

    “Spahn et al. (45) proposed that Tet(O) triggers the release of tetracycline through an allosteric mechanism.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning
    § RIBOSOMAL PROTECTION PROTEINS

    Protection of C1214 from DMS modification indicates that this base is directly shielded by Tet(O), supported by cryo-EM.

    “The protection of C1214 from DMS modification would indicate that this base is directly shielded by Tet(O), and this conclusion is supported by the fact that the Tet(O) binding site observed by cryo-EM approaches C1214 (Fig. 4B).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [12] 12. Connell, S. R., C. A. Trieber, U. Stelzl, E. Einfeldt, D. E. Taylor, and K. H. Nierhaus. 2002. The tetracycline resistance protein, Tet(O), perturbs the conformation of the ribosomal decoding center. Mol. Microbiol. 45: 14631472. source

      Not checked in the free run.

    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationnone given
    § RIBOSOMAL PROTECTION PROTEINS

    The discrepancy concerning the role of GTP hydrolysis probably does not reflect a functional difference but may result from different methods used in each laboratory.

    “This, however, probably does not reflect a functional difference, but instead, it may result from the different methods used in each laboratory.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    In the presence of tetracycline, the ribosome is blocked with an open A site, providing a kinetic window for Tet(O) to act and distinguishing the tetracycline-blocked ribosome from a translating ribosome.

    “In the presence of tetracycline, the ribosome is blocked with an open A site and this could provide a kinetic window for Tet(O) to act, thus distinguishing the tetracycline-blocked ribosome from a translating ribosome (11).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [11] 11. Connell, S. R., C. A. Trieber, E. Einfeldt, G. P. Dinos, D. E. Taylor, and K. Nierhaus. 2003. Mechanism of Tet(O)-mediated tetracycline resistance. EMBO J. 22: 945-953. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning
    § RIBOSOMAL PROTECTION PROTEINS

    Differences in domain IV may serve to distinguish Tet(O) and EF-G with respect to their activities; domain IV of EF-G more intimately overlaps with the A site-bound tRNA, consistent with its role in translocation, whereas interaction of domain IV of Tet(O) and h34 of the 30S subunit is consistent with its role in tetracycline release because h34 is a component of the primary tetracycline binding.

    “In this case, these differences in domain IV may serve to distinguish Tet(O) and EF-G with respect to their activities; namely, domain IV of EF-G more intimately overlaps with the A site-bound tRNA, an idea that is consistent with the role of domain IV of EF-G in translocation. In contrast, the interaction of domain IV of Tet(O) and h34 of the 30S subunit is consistent with its role in tetracycl”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

    • cites [4] 4. Brodersen, D. E., W. M. Clemons, A. P. Carter, R. J. Morgan-Warren, B. T. Wimberly, and V. Ramakrishnan. 2000. The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal subunit. Cell 103: 1143-1154. source

      Not checked in the free run.

    • cites [37] 37. Pioletti, M., F. Schlunzen, J. Harms, R. Zarivach, M. Gluhmann, H. Avila, A. Bashan, H. Bartels, T. Auerbach, C. Jacobi, T. Hartsch, A. Yonath, and F. Franceschi. 2001. Crystal structures of complexes of the small ribosomal subunit with tetracycline, edeine and IF3. EMBO J. 20: 1829-1839. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S1 (asserted). May combine two claims.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    Upon binding, Tet(O) contacts the base of h34, causing a disturbance in this helix that is propagated to the primary tetracycline-binding site, releasing the drug.

    “Spahn et al. (45) hypothesize that, upon binding, Tet(O) contacts the base of h34, which in turn causes a disturbance in this helix, which is propagated to the primary tetracycline-binding site, releasing the drug.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    The majority of interactions between Tet(O) and the ribosome are between Tet(O) and rRNA, with the only exception being a single interaction between domain III of Tet(O) and ribosomal protein S12.

    “In the cryo-EM study, Spahn et al. localize the sites of interaction between Tet(O) and the ribosome (Table 1), demonstrating that that the majority of the interactions are between Tet(O) and the rRNA (45). The only exception is a single interaction between domain III of Tet(O) and the ribosomal protein S12 (45).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    A direct interaction between Tet(O) and tetracycline bound to the primary site cannot be absolutely discounted, although chemical probing and cryo-EM suggest it is not the case.

    “It should be noted, however, that a direct interaction between Tet(O) and tetracycline bound to the primary site cannot be absolutely discounted, although both chemical probing and cryo-EM suggest this is not the case (12, 45).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [12] 12. Connell, S. R., C. A. Trieber, U. Stelzl, E. Einfeldt, D. E. Taylor, and K. H. Nierhaus. 2002. The tetracycline resistance protein, Tet(O), perturbs the conformation of the ribosomal decoding center. Mol. Microbiol. 45: 14631472. source

      Not checked in the free run.

    • cites [45] 45. Spahn, C. M. T., G. Blaha, R. K. Agrawal, P. Penczek, R. A. Grassucci, C. A. Trieber, S. R. Connell, D. E. Taylor, K. H. Nierhaus, and J. Frank. 2001. Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. Mol. Cell 7: 1037-1045. source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning
    § RIBOSOMAL PROTECTION PROTEINS

    Tet(O) and Tet(M) confer tetracycline resistance by releasing tetracycline from the ribosome and thereby freeing the ribosome from the inhibitory effects of the drug, such that aa-tRNA can bind to the A site and protein synthesis can continue.

    “Thus, it appears that Tet(O) and Tet(M) confer tetracycline resistance by releasing tetracycline from the ribosome and thereby freeing the ribosome from the inhibitory effects of the drug, such that aa-tRNA can bind to the A site and protein synthesis can continue.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    The enhancement of the C1402-C1501 cross-link may indicate that tetracycline promotes subtle structural rearrangement or fixes the ribosome in a conformation favorable for the cross-link.

    “These bases are distinct from the observed tetracycline binding sites and may indicate that tetracycline is promoting subtle structural rearrangement or fixing the ribosome in a conformation that is favorable for the establishment of the C1402-C1501 cross-link.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [32] 32. Noah, J. W., M. A. Dolan, P. Babin, and P. Wollenzien. 1999. Effects of tetracycline on the tertiary structure of ribosomal RNA in the Escherichia coli 30S ribosomal subunit. J. Biol. Chem. 274: 16576-16581. source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationreasoning
    § RIBOSOMAL PROTECTION PROTEINS

    h44 is a component of the decoding site and may have a role in the inhibitory action of tetracycline.

    “Additionally, as h44 is a component of the decoding site, it may have a role in the inhibitory action of tetracycline.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning
    § RIBOSOMAL PROTECTION PROTEINS

    The change might simply move the ribosome into a configuration compatible with stable tetracycline binding.

    “the change might simply move the ribosome into a configuration compatible with stable tetracycline binding.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationcitation
    § RIBOSOMAL PROTECTION PROTEINS

    The binding of tetracycline to the ribosome may be accompanied by a structural rearrangement.

    “it is possible that the binding of tetracycline to the ribosome (reaction f) is accompanied by a structural rearrangement.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation
    § RIBOSOMAL PROTECTION PROTEINS

    Tetracycline affects h44.

    “Noah et al. (32) presented evidence that tetracycline affects h44.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [32] 32. Noah, J. W., M. A. Dolan, P. Babin, and P. Wollenzien. 1999. Effects of tetracycline on the tertiary structure of ribosomal RNA in the Escherichia coli 30S ribosomal subunit. J. Biol. Chem. 274: 16576-16581. source

      Not checked in the free run.

25 of 25

  • The authors state this as an assumption.

    The other members of the RPP class [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through mechanisms similar to those of Tet(O) and Tet(M).

    “It is assumed, however, that the other members of this class of RPPs [Tet(S), Tet(T), Tet(Q), TetB(P), Tet(W), and OtrA] function through similar mechanisms.”

  • The paper relies on this without support.

    After being released, tetracycline is free to rebind the ribosome and again inhibit protein synthesis.

    “after being released, tetracycline is free to rebind the ribosome and again inhibit protein synthesis.”
2 of 2

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Tet(M) was initially described in Streptococcus spp. and Tet(O) in Campylobacter jejuni, but both are now widely distributed among bacterial species, likely because they are found in plasmids and broad-range conjugative transposons.

    “Tet(M) was initially described in Streptococcus spp. and Tet(O) in Campylobacter jejuni, but they are now both widely distributed among different bacterial species, likely because they have been found in several plasmids and in broad-range conjugative transposons (51).”

    From Mechanisms of Antibiotic Resistance (2016) · cites this paper as [51]

  • TetO was suggested to chase tetracycline from the ribosome indirectly via inducing local disturbances within h34.

    “As TetO is not observed to directly interact with C1054 or A1408 (14), TetO was suggested to chase tetracycline from the ribosome indirectly via inducing local disturbances within h34 (9,14,15).”

    From Structural basis for TetM-mediated tetracycline resistance (2012) · cites this paper as [9]

  • The conformational changes were proposed to persist after TetO has dissociated from the ribosome, preventing rebinding of tetracycline as well as stimulating delivery of the ternary complex.

    “Moreover, the conformational changes were proposed to persist after TetO has dissociated from the ribosome, preventing rebinding of tetracycline as well as stimulating delivery of the ternary complex (9,16).”

    From Structural basis for TetM-mediated tetracycline resistance (2012) · cites this paper as [9]

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Related

Authors
Connell, S. R., Tracz, D. M., Nierhaus, K. H., & Taylor, D. E.
Affiliations
  • University of Alberta (CA)
  • Max Planck Institute for Molecular Genetics (DE)
ORCID
Venue
Antimicrobial Agents and Chemotherapy
Publisher
American Society for Microbiology
Published
2003-12
Volume
47
Issue
12
Pages
3675-3681
ISSN
  • 0066-4804 (ISSN-L)
  • 1098-6596
DOI
10.1128/aac.47.12.3675-3681.2003
PMC
PMC296194
OpenAlex
W2168967909
Semantic Scholar
b51e42f14821598e8ac23c8bbd012170a3de473f
Cited by
449
References
60
Citations by year
2026: 22 · 2025: 35 · 2024: 20 · 2023: 18 · 2022: 19 · 2021: 28 · 2020: 20 · 2019: 29 · 2018: 31 · 2017: 28 · 2016: 23 · 2015: 21 · 2014: 23 · 2013: 17 · 2012: 17
FWCI
0.83
Type
journal article
Language
en
Open access
Licence
journals.asm.org/non-commercial-tdm-license (tdm)
Locations
  • Antimicrobial Agents and Chemotherapy (publishedVersion)
  • Europe PMC (PubMed Central) (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • Max Planck Institute for Plasma Physics (submittedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Medicine › Antibiotic Resistance in Bacteria
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA and protein synthesis mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords
ribosomal protection proteins, tetracycline resistance, tetM, tetO, ribosome protection, Campylobacter jejuni, Streptococcus, rRNA mutations, OTRA, tetW
MeSH terms
Bacteria, Bacterial Proteins, Models, Molecular, Ribosomes, Tetracycline Resistance

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Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-10.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-10.

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