Académique Documents
Professionnel Documents
Culture Documents
MINIREVIEW / MINISYNTHE`SE
Abstract: For more than half a century, tetracycline antibiotics have been used to treat infectious disease. However, what
once used to be a commonly prescribed family of antibiotics has now decreased in effectiveness due to wide-spread bacterial resistance. The chemical scaffold of the tetracyclines is a versatile and modifiable structure that is able to interact with
many cellular targets. The recent availability of detailed molecular interactions between tetracycline and its cellular targets,
along with an understanding of the tetracycline biosynthetic pathway, has provided us with a unique opportunity to usher
in a new era of rational drug design. Herein we discuss recent findings that have clarified the mode of action and the biosynthetic pathway of tetracyclines and that have shed light on the chemical biology of tetracycline antibiotics.
Key words: antibiotic, tetracycline, biosynthesis, resistance.
Resume : Depuis plus dun demi-sie`cle, les antibiotiques de la famille des tetracyclines ont ete utilises pour traiter les maladies infectieuses. Cependant, la prescription generalisee de cette famille dantibiotiques en a diminue lefficacite a` cause
dune resistance bacterienne repandue. Lechafaudage chimique des tetracyclines est une structure versatile et modifiable
qui est capable dinteragir avec plusieurs cibles cellulaires. La recente disponibilite de donnees detaillees des interactions
moleculaires de la tetracycline et ses cibles cellulaires conjuguee a` une meilleure comprehension de la voie de biosynthe`se
de la tetracycline nous a fourni une opportunite unique dinaugurer une nouvelle e`re de conception rationnelle de medicaments. Nous discutons ci-dessous des recentes decouvertes qui ont permis de clarifier le mode daction et la voie de biosynthe`se des tetracyclines, et qui ont fait la lumie`re sur la chimie biologique des antibiotiques de la famille des
tetracyclines.
Mots-cles : antibiotique, tetracycline, biosynthe`se, resistance.
[Traduit par la Redaction]
Introduction
Tetracyclines (Fig. 1) were first discovered in 1948 by
Benjamin Duggar (Duggar 1948) as natural products produced by species of Streptomyces, and have since proven to
be an economically valuable drug class over the past 6 decades. The highly modified chemical scaffold of the tetracyclines affords them versatility and allows them to interact
with a variety of biological targets. A remarkable feature of
tetracyclines is the presence of keto-enol functional groups
on one face of the scaffold, providing them with the ability
to chelate divalent cations, which plays a prominent role in
their biological functions. Although the chemical biology of
tetracycline antibiotics has been extensively studied, there
Received 15 October 2007. Revision received 12 December
2007. Accepted 19 December 2007. Published on the NRC
Research Press Web site at bcb.nrc.ca on 27 March 2008.
B. Zakeri and G.D. Wright.2 Department of Biochemistry and
Biomedical Sciences, DeGroote School of Medicine, McMaster
University, 1200 Main St. W, Hamilton, ON L8N 3Z5, Canada.
1This
doi:10.1139/O08-002
125
rial resistance and a corresponding decline in the use of tetracycline antibiotics for human therapy.
Recently many advances have been made in elucidating
the structureactivity relationships, resistance mechanisms,
and biosynthetic pathways of tetracyclines. The availability
of high resolution crystallographic structures of tetracycline
bound to the 30S ribosomal subunit and a detailed understanding of the tailoring steps in the biosynthetic pathways
will result in exciting new strategies for the rational design
of novel tetracycline derivatives. Herein we review new
findings that shed light on how tetracyclines interact with
their cellular targets and the secondary metabolic pathways
that lead to their production from simple precursor molecules.
Tetracycline antibiotics
The first members of the tetracycline antibiotics to be described, chlortetracycline (CTC) and oxytetracycline (OTC)
(Fig. 1), were discovered in the late 1940s during the early
period of the Golden Age of antibiotic discovery as a result
126
clines, both in the clinic and in agriculture, imposed a massive selection pressure for resistant isolates. Not surprisingly, by 1953 the first tetracycline-resistant strains of
Shigella dysenteriae were isolated, and soon afterwards in
1955, multidrug resistant Shigella species that were resistant
to tetracycline, chloramphenicol, and streptomycin were discovered (Watanabe 1963; Roberts 1996).
To keep pace with the rapid emergence of resistant pathogens, new tetracyclines were sought out; however, new tetracyclines from traditional microbial sources proved
difficult to find. As a result, we entered a golden age of
antibiotic medicinal chemistry, during which, scientists
tried to improve potency and evade resistance mechanisms
by chemically modifying antibiotic scaffolds (Wright 2007).
This resulted in the development of semisynthetic secondgeneration tetracyclines, which include doxycycline and
minocycline (Fig. 1). These compounds are more lipophilic
than their parent compounds, and as a result, they have better absorption and pharmacokinetic parameters (Agwuh and
MacGowan 2006). Recently, third-generation tetracyclines,
which include the semisynthetic glycylcyclines and aminomethylcyclines, have been developed (Sum and Petersen
1999; Agwuh and MacGowan 2006), some of which are
currently in clinical trials (e.g., Paratek Pharmaceuticals,
www.paratekpharm.com/pt_tet_inhib.html). Tigecycline is a
member of the glycylcyclines, and was approved for clinical
use by the US Food and Drug Administration in 2005. It
contains an N-alkyl-glycylamido group at position 9 on the
scaffold of minocycline (Fig. 1). This modification results
in a more efficient interaction with the ribosome and the
evasion of classic tetracycline-resistance mechanisms such
as efflux pumps and ribosomal protection proteins (vide infra) (Slover et al. 2007).
Tetracyclines and indeed, many other natural products,
often possess very complex chemical structures that are produced by a large number of enzymes in a "conveyor-belt"
fashion. Once the scaffolds of these compounds are produced, many regio-specific tailoring reactions position functional groups at strategic location, allowing the compounds
to interact with biological targets via hydrogen bonding. As
a result, tetracyclines contain several chiral centres and
many chemically reactive substituents, which makes their
total chemical synthesis challenging. Furthermore, their
chemical sensitivity to acidic and basic media, which cause
the molecules to undergo chemical transformations, further
hinders synthetic approaches (Muxfeldt et al. 1979). For
these reasons, it would seem that the ideal method of developing novel tetracycline derivatives would be through semisynthetic approaches. Nevertheless, successful attempts at
the total synthesis of tetracyclines have been made. In 1979,
Muxfeldt et al. (Muxfeldt et al. 1979) reported the total synthesis of OTC from a tetralone aldehyde producing 47 other
precursor molecules before isolating OTC in an unspecified
yield. Further improvements to this approach have been
made recently by Myers group. In 2005 they reported the
synthesis of ()-tetracycline from benzoic acid in 17 steps
with an overall yield of 1.1% (Charest et al. 2005a). In the
same year, they also reported the synthesis of ()-doxycycline, also from benzoic acid, with an 8.3% yield over 18
steps (Charest et al. 2005b). However, total syntheses of
such complex molecules are time consuming, labour inten-
Mode of action
Tetracyclines impart their broad spectrum antibiotic activities through both bacteriostatic and bactericidal modes of
action. The clinically used compounds are bacteriostatic and
their mode of action has been well characterized, whereas
the action of bactericidal tetracyclines, such as chelocardin
(Fig. 1), is poorly understood (Schnappinger and Hillen
1996). Recently, Kohanski et al. have shown that the cellular killing capabilities of three major classes of bactericidal
antibiotics (quinolones, b-lactams, and aminoglycosides) can
be partly credited to the production of toxic hydroxyl radicals (Kohanski et al. 2007). Although there is no evidence
of hydroxyl radical formation by bactericidal tetracyclines,
it does warrant further investigation. The antimicrobial function of tetracycline has been attributed to the binding of the
bacterial 30S ribosomal subunit near the A site and subsequent inhibition of protein synthesis by the prevention of
aminoacylated-tRNA docking (Maxwell 1967; Brodersen et
al. 2000). However, there is considerable biochemical evidence for the presence of one strong and several minor binding sites for tetracycline in the ribosome, which has led to
discrepancies in the mode of action and the functions of the
minor binding sites reported in the literature (Tritton 1977;
Epe and Woolley 1984). The recent revelation of the molecular interactions between tetracycline and the ribosome
through X-ray crystallography has clarified their mode of
action.
Uptake
To gain access to the ribosome, tetracyclines must first
traverse biological membranes and do so in both gram-positive and gram-negative organisms. It has long been known
that tetracyclines chelate divalent cations, commonly mag#
127
Tet-2, which binds in a hydrophobic pocket made by a ribosomal protein (Pioletti et al. 2001). Pioletti et al. proposed that these sites may function synergistically with the
Tet-1 to account for the protein inhibitory action of tetracyclines (Pioletti et al. 2001). All of the binding sites, except for Tet-3, contact rRNA binding proteins that are
required for the assembly of new 30S subunits, and thus
tetracycline molecule binding may disrupt this process and
inhibit the early steps of protein biosynthesis (Auerbach et
al. 2002; Zarivach et al. 2002). Of particular interest is the
Tet-5 site, where tetracycline makes hydrophilic contacts
with H27 and H11 and is enclosed in a tight pocket surrounded by H20, H27, and S17 (Pioletti et al. 2001). This
site, which was also observed in the Brodersen et al.
model, is interesting because the switch helix H27 is located in the decoding centre of the 16S RNA. Along with
H44, H27 undergoes conformational changes to accommodate base pairing interactions between tRNA anti-codons
and mRNA codons, which are pivotal interactions in the
decoding process. Tetracycline binding to Tet-5 may limit
these conformation changes, which leads to the disruption
of translation, and hence, the inhibition of protein synthesis
(Auerbach et al. 2002; Zarivach et al. 2002). These detailed accounts of molecular interactions and the location
of the Tet-1 site shed light on the chemical diversity that
is present in the tetracycline group of antibiotics, in which
variations are restricted to the hydrophobic side of the
molecules.
Analysis of the topography of the Tet-1 site in the presence of tetracycline provides insight into the chemical interactions between tetracyclines and the ribosome (Fig. 2b).
The hydrophilic side of the molecule is responsible for the
majority of the chemical interactions with the 16S RNA
moiety of the ribosome, whereas the more hydrophobic side
appears to be in more open space. This is consistent with
chemical modifications on the hydrophobic side that have
been shown to be well tolerated by tetracyclines while still
maintaining their cell-growth inhibitory properties. Modifications made to the tetracycline scaffold on C1, C10, C11,
and C12 (Fig. 2a) lead to a loss of antimicrobial activity
(Nelson et al. 1994). On the other hand, modifications can
be made in positions C2, C4, C5, C6, C7, C8, and C9 while
still maintaining the protein synthesis inhibitory activity of
the parent compound, as can be seen among the natural
product compounds of the tetracycline family (Fig. 1)
(Nelson 2002). This molecular model is in agreement with
the bioactive semisynthetic tetracycline derivatives that
have been made. Modifications have been made to tetracycline scaffolds to improve their pharmacokinetic properties,
evade clinically relevant resistance mechanisms, improve
antimicrobial potency, and modify the compounds for medicinal uses other than antibiotics. However, with our current understanding, there is potential for a new phase of
tetracycline design in which analogues can be rationally developed by first modelling them into the ribosomal binding
site, and then modifying them in an attempt to disrupt key
interactions involved in resistance.
Atypical tetracyclines
Certain tetracycline analogues, such as chelocardin and 6thiatetracycline, are termed atypical tetracyclines because
#
128
Fig. 2. (a) A labelled tetracycline compound in complex with Mg2+. The highlighted region represents substituents that are required for the
protein inhibitory activity of tetracyclines (Nelson et al. 1994; Nelson 2002). (b) Tetracycline in complex with the 30S ribosomal subunit of
Thermus thermophilus, as determined by Pioletti et al. (PDB file 1I97) (Pioletti et al. 2001). The magnified region in the left panel illustrates tetracycline in complex with Mg2+ (yellow sphere) bound in the Tet-1 binding pocket of the 30S ribosomal subunit. The magnified
region in the right panel illustrates a green tetracycline molecule in complex with a yellow Mg2+ ion bound to the Tet-1 binding pocket of
the 30S ribosomal subunit of T. thermophilus, and a red nucleotide on the left of the image, which corresponds to the equivalent position of
base 1058 in the 16S rRNA sequence of Escherichia coli. In propionibacteria, a mutation from cytosine to guanine at base 1058 leads to
tetracycline resistance (Ross et al. 1998).
1991). Furthermore, when added to growing bacterial cultures, tetracyclines were shown to limit culture growth by
inhibiting the incorporation of amino acids in protein synthesis. However, atypical tetracyclines not only inhibit protein synthesis but also completely prevent the incorporation
of nucleic acids into DNA and RNA, which led to the theory
that these molecules function by disrupting the cytoplasmic
membrane (Rasmussen et al. 1991). Further evidence for
this notion was provided when it was shown that, in the
presence of atypical tetracyclines, E. coli cytoplasmic membranes undergo morphological alterations that lead to eventual cell lysis (Oliva et al. 1992). In solution, tetracyclines
exist as an equilibrium mixture of two species, in which
#
one form is lipophilic and non-ionized while the other is hydrophilic and zwitterionic (Hughes et al. 1979). The lipophilic form is important during drug uptake, whereas the
hydrophilic form is required for ribosomal binding. The preferred species of chelocardin, and presumably of the other
atypical tetracyclines, is apparently the lipophilic form,
which along with the planarity of the BCD rings in the molecules, provides the ideal structure for the compounds to enter the hydrophobic cytoplasmic membrane. They are likely
to be retained once they are in the membrane, where they
cause membrane damage and eventual lysis (Chopra et al.
1992; Chopra 1994; Rasmussen et al. 1991). This may explain the adverse side effects seen with the therapeutic use
of these compounds, which may be because of indiscriminate interactions with both prokaryotic and eukaryotic membranes (Chopra et al. 1992). This possible mode of action is
in agreement with the observation that the atypical tetracyclines retain their antibiotic activities even in the presence
of the two most common forms tetracycline resistance, drug
efflux and ribosomal protection (Oliva and Chopra 1992).
The protein synthesis inhibitory action of tetracyclines,
combined with a versatile structure that allows them to traverse biological membranes with ease, are key to the broad
spectrum activity of this group of antibiotics. Consequently,
their extensive use has lead to the emergence of wide-spread
resistance among both pathogenic and nonpathogenic bacteria, which has severely crippled the clinical application of
these drugs.
Resistance
Genes that confer tetracycline resistance are among some
of the most commonly found antibiotic resistance determinants in bacteria. They are often located on mobile genetic
elements, such as plasmids and transposons, which can be
passed among bacteria of different genera through horizontal
gene transfer (Speer et al. 1992). Also, common resistance
mechanisms are nondestructive, leading to a prolonged exposure of bacteria to tetracyclines that results in an extended
period of selection for resistant isolates (Hillen and Berens
1994). The nomenclature, mechanisms, and epidemiology of
tetracycline resistance has been extensively reviewed elsewhere (Refer to references Connell et al. 2003; Speer et al.
1992; Roberts 2003, 2005; Chopra and Roberts 2001) and
will therefore be only briefly discussed here.
There are four mechanisms by which bacteria become resistant to tetracyclines (Fig. 3): (i) by reducing the intracellular concentration of the compound through active efflux
(Ball et al. 1980), (ii) through disruption of the tetracyclineribosomal interaction by ribosomal protection proteins
(RPPs) (Burdett 1991), (iii) by enzymatic inactivation of the
drug through monohydroxylation (Yang et al. 2004), and
(iv) by altering the target site through 16S RNA mutation
(Ross et al. 1998). Of these, tetracycline efflux proteins and
ribosomal protection proteins are by far the most common
forms of resistance encountered among pathogenic and environmental bacteria (Chopra and Roberts 2001). As of July
2007, there were 40 identified tetracycline resistance genes,
designated tet and otr genes. Of these, 25 encode efflux
proteins, 11 encode ribosomal protection proteins, 3 are
tetracycline-degrading enzymes, and 1 has an unknown
129
Fig. 3. Tetracycline resistance mechanisms.
130
Tetracycline biosynthesis
Microbes invest large amounts of cellular energy for the
synthesis of an array of secondary metabolites with varying
degrees of complexity, which have been used to treat many
human ailments. Natural products have a wide variety of biological functions to aid the host organism, including their
highly exploited antimicrobial activity, hormone-like regulation of differentiation, quorum sensing, roles in metal transport, and diffusible siderophores (Challis and Hopwood
2003; Linares et al. 2006). These activities are critical to
the survival of the host organisms in the harsh and competitive environments in which they live. It is therefore important to understand how these molecules are made so that we
can take advantage of the efficiency of biological systems to
increase the chemical diversity that is available to us.
The biosynthesis of CTC and OTC, the first two tetracyclines discovered, was first studied in the early 1960s
through blocked mutant analysis, substrate feeding
experiments, and precursor analysis (Miller et al. 1964;
McCormick et al. 1965, 1968; McCormick and Jensen
1965, 1969; Barbatschi et al. 1965).
However, it was not until recently that we have come to
better understand the mechanisms by which these two compounds are made and the genetic machinery involved in the
process. The biosynthetic gene clusters that encode CTC
(Ryan 1999; Nakano et al. 2004) and OTC (Zhang et al.
2006) synthesis have been sequenced and have provided an
insight into tetracycline biosynthesis. The genetic structure
and enzymatic mechanisms for the biosynthesis of the two
antibiotics are very similar and therefore only OTC biosynthesis will be discussed henceforth.
Streptomyces rimosus, the OTC producer, has an 8 Mb
linear chromosome with the OTC biosynthetic cluster situated approximately 600 kb from one end of the chromosome
which contributes to its genetic instability (Pandza et al.
#
131
132
Fig. 4. A summary of oxytetracycline biosynthesis in Streptomyces rimosus. (a) The oxytetracycline biosynthetic gene cluster, as determined
by Zhang et al. (2006). (b) An updated model, proposed by Zhang et al. (2006), of the oxytetracycline biosynthetic pathway.
should not affect the function of the minimal PKS, has led
to the production of novel truncated polyketide molecules.
OxyK catalyzes ring closure of the full length polyketide
chain, implying that it acts upon the polyketide after the
minimal PKS has completed the chain. Yet, its disruption
led to the production of four novel polyketides with shorter
chain length compared with that in OTC (Petkovic et al.
1999). Furthermore, disruption of oxyS, which encodes an
anhydrotetracycline oxygenase that hydroxylates C6 late in
the OTC biosynthetic pathway, also led to the production of
a truncated polyketide (Peric-Concha et al. 2005). These results imply that the minimal PKS (KSa, KSb, and ACP) may
not constitute a self-sufficient enzyme complex and that it
may require other enzymes in the OTC biosynthetic pathway
for complete activity (Peric-Concha et al. 2005). The problems encountered with gene deletions in S. rimosus, compounded by the challenges associated with heterologous
expression of biosynthetic proteins and a lack of availability
Future directions
Antibiotics are one of the most successful classes of drugs
that have ever been used in human therapy. They represent a
25 billion $US annual market, which is expected to steadily
increase (Christoffersen 2006). So why are big pharmaceutical companies shutting down their antibiotic discovery programs? Although there has been a rise in multidrug resistant
pathogens and there is a desperate need for new antibiotics,
the rewards are not substantial enough to overcome the
#
133
Fig. 5. The structure of COL-3, a matrix metalloproteinase inhibitor
vides us with the necessary tools to rationally design semisynthetic compounds to combat emerging multidrug
resistant pathogens. The broad-spectrum antibiotic activity
of tetracyclines makes them a vital weapon against infectious disease and, as evidenced by the development of tigecycline, they remain important scaffolds for new chemists.
Acknowledgement
Tetracycline research in our laboratory is funded by the
Natural Sciences and Engineering Research Council of Canada.
References
Acharya, M.R., Venitz, J., Figg, W.D., and Sparreboom, A. 2004.
Chemically modified tetracyclines as inhibitors of matrix metalloproteinases. Drug Resist. Updat. 7: 195208. doi:10.1016/j.
drup.2004.04.002. PMID:15296861.
Agwuh, K.N., and MacGowan, A. 2006. Pharmacokinetics and
pharmacodynamics of the tetracyclines including glycylcyclines.
J. Antimicrob. Chemother. 58: 256265. doi:10.1093/jac/dkl224.
PMID:16816396.
Auerbach, T., Bashan, A., Harms, J., Schluenzen, F., Zarivach, R.,
Bartels, H., et al. 2002. Antibiotics targeting ribosomes: crystallographic studies. Curr. Drug Targets Infect. Disord. 2: 169186.
doi:10.2174/1568005023342506. PMID:12462147.
Ball, P.R., Shales, S.W., and Chopra, I. 1980. Plasmid-mediated
tetracycline resistance in Escherichia coli involves increased efflux of the antibiotic. Biochem. Biophys. Res. Commun. 93: 74
81. doi:10.1016/S0006-291X(80)80247-6. PMID:6990931.
Baltz, R.H. 2005. Antibiotic discovery from actinomycetes: Will a
renaissance follow the decline and fall? SIM News, 55: 186
196.
Barbatschi, F., Dann, M., Martin, J.H., Miller, P., Mitscher, L.A.,
and Bohonos, N. 1965. 4-Dedimethylamino-4-epiamino-5a,6-anhydrotetracycline. Experientia, 21: 162163. doi:10.1007/
BF02141995. PMID:5830054.
Brodersen, D.E., Clemons, W.M., Jr., Carter, A.P., MorganWarren, R.J., Wimberly, B.T., and Ramakrishnan, V. 2000.
The structural basis for the action of the antibiotics tetracycline, pactamycin, and hygromycin B on the 30S ribosomal
subunit. Cell, 103: 11431154. doi:10.1016/S0092-8674(00)
00216-6. PMID: 11163189.
Burdett, V. 1991. Purification and characterization of Tet(M), a
protein that renders ribosomes resistant to tetracycline. J. Biol.
Chem. 266: 28722877. PMID:1993661.
Burdett, V. 1996. Tet(M)-promoted release of tetracycline from
ribosomes is GTP dependent. J. Bacteriol. 178: 32463251.
PMID:8655505.
#
134
Challis, G.L., and Hopwood, D.A. 2003. Synergy and contingency
as driving forces for the evolution of multiple secondary metabolite production by Streptomyces species. Proc. Natl. Acad.
Sci. U.S.A. 100(Suppl 2): 1455514561. doi:10.1073/pnas.
1934677100. PMID:12970466.
Charest, M.G., Siegel, D.R., and Myers, A.G. 2005a. Synthesis of
()-tetracycline. J. Am. Chem. Soc. 127: 82928293. doi:10.
1021/ja052151d. PMID:15941256.
Charest, M.G., Lerner, C.D., Brubaker, J.D., Siegel, D.R.,
and Myers, A.G. 2005b. A Convergent enantioselective
route to structurally diverse 6-deoxytetracycline antibiotics.
Science, 308: 395398. doi:10.1126/science.1109755. PMID:
15831754.
Chopra, I. 1994. Tetracycline analogs whose primary target is not
the bacterial ribosome. Antimicrob. Agents Chemother. 38:
637640. PMID:8031024.
Chopra, I., and Roberts, M. 2001. Tetracycline antibiotics: mode of
action, applications, molecular biology, and epidemiology of
bacterial resistance. Microbiol. Mol. Biol. Rev. 65: 232260.
doi:10.1128/MMBR.65.2.232-260.2001. PMID:11381101.
Chopra, I., Hawkey, P.M., and Hinton, M. 1992. Tetracyclines, molecular and clinical aspects. J. Antimicrob. Chemother. 29: 245
277. doi:10.1093/jac/29.3.245. PMID:1592696.
Christoffersen, R.E. 2006. Antibioticsan investment worth making? Nat. Biotechnol. 24: 15121514. doi:10.1038/nbt12061512. PMID:17160052.
Connell, S.R., Tracz, D.M., Nierhaus, K.H., and Taylor, D.E. 2003.
Ribosomal protection proteins and their mechanism of tetracycline resistance. Antimicrob. Agents Chemother. 47: 36753681.
doi:10.1128/AAC.47.12.3675-3681.2003. PMID:14638464.
Cundliffe, E. 1989. How antibiotic-producing organisms avoid suicide. Annu. Rev. Microbiol. 43: 207233. doi:10.1146/annurev.
mi.43.100189.001231. PMID:2679354.
DCosta, V.M., McGrann, K.M., Hughes, D.W., and Wright, G.D.
2006. Sampling the antibiotic resistome. Science, 311: 374377.
doi:10.1126/science.1120800. PMID:16424339.
Davies, J. 1994. Inactivation of antibiotics and the dissemination of
resistance genes. Science, 264: 375382. doi:10.1126/science.
8153624. PMID:8153624.
Degenkolb, J., Takahashi, M., Ellestad, G.A., and Hillen, W. 1991.
Structural requirements of tetracyclineTet repressor interaction:
determination of equilibrium binding constants for tetracycline
analogs with the Tet repressor. Antimicrob. Agents Chemother.
35: 15911595. PMID:1929330.
Duggar, B.M. 1948. Aureomycin: a product of the continuing
search for new antibiotics. Ann. N. Y. Acad. Sci. 51: 177181.
doi:10.1111/j.1749-6632.1948.tb27262.x. PMID:18112227.
Eckert, B., and Beck, C.F. 1989. Overproduction of transposon
Tn10-encoded tetracycline resistance protein results in cell death
and loss of membrane potential. J. Bacteriol. 171: 35573559.
PMID:2542231.
Epe, B., and Woolley, P. 1984. The binding of 6-demethylchlortetracycline to 70S, 50S and 30S ribosomal particles: a quantitative study by fluorescence anisotropy. EMBO J. 3: 121126.
PMID:6423382.
Fernandes, P. 2006. Antibacterial discovery and developmentthe
failure of success? Nat. Biotechnol. 24: 14971503. doi:10.1038/
nbt1206-1497. PMID:17160049.
Guiney, D.G., Jr., Hasegawa, P., and Davis, C.E. 1984. Expression
in Escherichia coli of cryptic tetracycline resistance genes from
Bacteroides R plasmids. Plasmid, 11: 248252. doi:10.1016/
0147-619X(84)90031-3. PMID:6379711.
Hertweck, C., Luzhetskyy, A., Rebets, Y., and Bechthold, A. 2007.
Type II polyketide synthases: gaining a deeper insight into enzy-
135
novel polyketides with altered chain length. J. Biol. Chem. 280:
3745537460. doi:10.1074/jbc.M503191200. PMID:16148009.
Petkovic, H., Thamchaipenet, A., Zhou, L.H., Hranueli, D., Raspor,
P., Waterman, P.G., and Hunter, I.S. 1999. Disruption of an aromatase/cyclase from the oxytetracycline gene cluster of Streptomyces rimosus results in production of novel polyketides with
shorter chain lengths. J. Biol. Chem. 274: 3282932834.
PMID:10551844.
Pioletti, M., Schlunzen, F., Harms, J., Zarivach, R., Gluhmann, M.,
Avila, H., et al. 2001. Crystal structures of complexes of the
small ribosomal subunit with tetracycline, edeine and IF3.
EMBO J. 20: 18291839. doi:10.1093/emboj/20.8.1829. PMID:
11296217.
Poole, K. 2005. Efflux-mediated antimicrobial resistance. J. Antimicrob. Chemother. 56: 2051. doi:10.1093/jac/dki171. PMID:
15914491.
Poole, K. 2007. Efflux pumps as antimicrobial resistance mechanisms. Ann. Med. 39: 162176. doi:10.1080/07853890701195262.
PMID:17457715.
Ramos, J.L., Martinez-Bueno, M., Molina-Henares, A.J., Teran,
W., Watanabe, K., Zhang, X., et al. 2005. The TetR family of
transcriptional repressors. Microbiol. Mol. Biol. Rev. 69: 326
356. doi:10.1128/MMBR.69.2.326-356.2005. PMID:15944459.
Rasmussen, B., Noller, H.F., Daubresse, G., Oliva, B., Misulovin,
Z., Rothstein, D.M., et al. 1991. Molecular basis of tetracycline
action: identification of analogs whose primary target is not the
bacterial ribosome. Antimicrob. Agents Chemother. 35: 2306
2311. PMID:1725100.
Roberts, M.C. 1996. Tetracycline resistance determinants: mechanisms of action, regulation of expression, genetic mobility, and
distribution. FEMS Microbiol. Rev. 19: 124. doi:10.1111/j.
1574-6976.1996.tb00251.x. PMID:8916553.
Roberts, M.C. 2003. Tetracycline therapy: update. Clin. Infect. Dis.
36: 462467. doi:10.1086/367622. PMID:12567304.
Roberts, M.C. 2005. Update on acquired tetracycline resistance
genes. FEMS Microbiol. Lett. 245: 195203. doi:10.1016/j.
femsle.2005.02.034. PMID:15837373.
Roberts, M.C. 2007. Mechanism of resistance for characterized tet
and otr genes [online]. Available from http://faculty.washington.
edu/marilynr/tetweb1.pdf [cited August 2007].
Ross, J.I., Eady, E.A., Cove, J.H., and Cunliffe, W.J. 1998. 16S
rRNA mutation associated with tetracycline resistance in a
gram-positive bacterium. Antimicrob. Agents Chemother. 42:
17021705. PMID:9661007.
Ryan, M.J. 1999. Strain for the production of 6-dimethyltetracycline, method for producing the strain and vector for use in the
method. US Patent 5989903.
Sapadin, A.N., and Fleischmajer, R. 2006. Tetracyclines: nonantibiotic properties and their clinical implications. J. Am. Acad.
Dermatol.
54:
258265.
doi:10.1016/j.jaad.2005.10.004.
PMID:16443056.
Schnappinger, D., and Hillen, W. 1996. Tetracyclines: antibiotic
action, uptake, and resistance mechanisms. Arch. Microbiol.
165: 359369. doi:10.1007/s002030050339. PMID:8661929.
Slover, C.M., Rodvold, K.A., and Danziger, L.H. 2007. Tigecycline: a novel broad-spectrum antimicrobial. Ann. Pharmacother.
41: 965972. doi:10.1345/aph.1H543. PMID:17519296.
Spahn, C.M., Blaha, G., Agrawal, R.K., Penczek, P., Grassucci,
R.A., Trieber, C.A., et al. 2001. Localization of the ribosomal
protection protein Tet(O) on the ribosome and the mechanism
of tetracycline resistance. Mol. Cell, 7: 10371045. doi:10.
1016/S1097-2765(01)00238-6. PMID:11389850.
Speer, B.S., and Salyers, A.A. 1988. Characterization of a novel
tetracycline resistance that functions only in aerobically
#
136
grown Escherichia coli. J. Bacteriol. 170: 14231429. PMID:
2832361.
Speer, B.S., Shoemaker, N.B., and Salyers, A.A. 1992. Bacterial resistance to tetracycline: mechanisms, transfer, and clinical significance. Clin. Microbiol. Rev. 5: 387399. PMID:1423217.
Stokstad, E.L.R., Jukes, T.H., Pierce, J., Page, A.C., Jr., and
Franklin, A.L. 1949. The multiple nature of the animal protein
factor. J. Biol. Chem. 180: 647654. PMID:18135798.
Sum, P.E., and Petersen, P. 1999. Synthesis and structure-activity
relationship of novel glycylcycline derivatives leading to the discovery of Gar-936. Bioorg. Med. Chem. Lett. 9: 14591462.
doi:10.1016/S0960-894X(99)00216-4. PMID:10360756.
Takahashi, M., Degenkolb, J., and Hillen, W. 1991. Determination
of the equilibrium association constant between Tet repressor
and tetracycline at limiting Mg2+ concentrations: a generally applicable method for effector-dependent high-affinity complexes.
Anal. Biochem. 199: 197202. doi:10.1016/0003-2697(91)
90089-C. PMID:1812784.
Tritton, T.R. 1977. Ribosome-tetracycline interactions. Biochemistry, 16: 41334138. doi:10.1021/bi00637a029. PMID:334241.
Watanabe, T. 1963. Infective heredity of multiple drug resistance
in bacteria. Bacteriol. Rev. 27: 87115. PMID:13999115.
White, J.P., and Cantor, C.R. 1971. Role of magnesium in the binding of tetracycline to Escherichia coli ribosomes. J. Mol. Biol.