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The Crystal Structure of Two Macrolide Glycosyltransferases Provides a Blueprint for Host Cell Antibiotic Immunity

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The Crystal Structure of Two Macrolide Glycosyltransferases Provides a Blueprint for Host Cell Antibiotic Immunity Corrections BIOCHEMISTRY. For the article ‘‘The crystal structure of two GENETICS. For the article ‘‘Deletion of the orphan nuclear recep- macrolide glycosyltransferases provides a blueprint for host cell tor COUP-TFII in uterus leads to placental deficiency,’’ by antibiotic immunity,’’ by David N. Bolam, Shirley Roberts, Mark Fabrice G. Petit, Soazik P. Jamin, Isao Kurihara, Richard R. R. Proctor, Johan P. Turkenburg, Eleanor J. Dodson, Carlos Behringer, Francesco J. DeMayo, Ming-Jer Tsai, and Sophia Y. Martinez-Fleites, Min Yang, Benjamin G. Davis, Gideon J. Tsai, which appeared in issue 15, April 10, 2007, of Proc Natl Davies, and Harry J. Gilbert, which appeared in issue 13, March Acad Sci USA (104:6293–6298; first published April 2, 2007; 27, 2007, of Proc Natl Acad Sci USA (104:5336–5341; first 10.1073͞pnas.0702039104), the authors note that the e-mail published March 21, 2007; 10.1073͞pnas.0607897104), the au- address for corresponding author Fabrice G. Petit appeared thors note that, in addition to writing the paper, Benjamin G. incorrectly. The correct address is [email protected]. The Davis should be credited with designing the research and online version has been corrected. analyzing the data. The corrected author contributions footnote ͞ ͞ ͞ ͞ appears below. www.pnas.org cgi doi 10.1073 pnas.0703353104 Author contributions: D.N.B., S.R., and M.R.P. contributed equally to this work; D.N.B., M.R.P., B.G.D., G.J.D., and H.J.G. PHYSIOLOGY. For the article ‘‘Night eating and obesity in the designed research; D.N.B., S.R., and M.R.P. performed re- EP3R-deficient mouse,’’ by Manuel Sanchez-Alavez, Izabella search; D.N.B., M.R.P., J.P.T., E.J.D., C.M.-F., M.Y., B.G.D., Klein, Sara E. Brownell, Iustin V. Tabarean, Christopher N. and H.J.G. analyzed data; and B.G.D., G.J.D., and H.J.G. wrote Davis, Bruno Conti, and Tamas Bartfai, which appeared in issue the paper. 8, February 20, 2007, of Proc Natl Acad Sci USA (104:3009–3014; first published February 16, 2007; 10.1073͞pnas.0611209104), ͞ ͞ ͞ ͞ www.pnas.org cgi doi 10.1073 pnas.0704090104 the authors note that the following acknowledgment was inad- vertently omitted from the article: ‘‘We thank Professor Shuh Narumiya (Kyoto University, Kyoto, Japan) (4, 5, 21) for the ENVIRONMENTAL SCIENCES. For the article ‘‘Combined climate and generation and subsequent generous transfer of the EP3RϪ/Ϫ carbon-cycle effects of large-scale deforestation,’’ by G. Bala, K. mice.’’ Caldeira, M. Wickett, T. J. Phillips, D. B. Lobell, C. Delire, and A. Mirin, which appeared in issue 16, April 17, 2007, of Proc Natl www.pnas.org͞cgi͞doi͞10.1073͞pnas.0704094104 Acad Sci USA (104:6550–6555; first published April 9, 2007; 10.1073͞pnas.0608998104), the authors note the following. On page 6550, left column, in paragraph 1 of the main text, line 7, the phrase ‘‘However, deforestation also exerts a cooling influ- ence by (iv) decreasing the surface albedo’’ should instead read: ‘‘However, deforestation also exerts a cooling influence by (iv) increasing the surface albedo.’’ Also on page 6550, right column, in Results, paragraph 2, line 1, the phrase ‘‘the atmospheric CO2 concentration is higher by 299, 110, and 5 ppmv’’ should instead read: ‘‘the atmospheric CO2 concentration is higher by 199, 110, and 5 ppmv.’’ Lastly, in the Fig. 1 legend, line 8, the phrase ‘‘net cooling, near-zero temperature change, and net warming, re- spectively’’ should instead read: ‘‘net warming, near-zero tem- perature change, and net cooling, respectively.’’ These errors do not affect the conclusions of the article. CORRECTIONS www.pnas.org͞cgi͞doi͞10.1073͞pnas.0704096104 www.pnas.org PNAS ͉ June 5, 2007 ͉ vol. 104 ͉ no. 23 ͉ 9911 Downloaded by guest on September 28, 2021 The crystal structure of two macrolide glycosyltransferases provides a blueprint for host cell antibiotic immunity David N. Bolam*, Shirley Roberts†, Mark R. Proctor*, Johan P. Turkenburg†, Eleanor J. Dodson†, Carlos Martinez-Fleites†, Min Yang‡, Benjamin G. Davis‡, Gideon J. Davies†§, and Harry J. Gilbert*§ *Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle upon Tyne, Framlington Place, Newcastle upon Tyne NE2 4HH, United Kingdom; †Structural Biology Laboratory, Department of Chemistry, University of York, Heslington, York YO10 5YW, United Kingdom; and ‡Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Mansfield Road, Oxford OX1 3TA, United Kingdom Edited by Jon Clardy, Harvard Medical School, Boston, MA, and accepted by the Editorial Board January 27, 2007 (received for review September 11, 2006) Glycosylation of macrolide antibiotics confers host cell immunity from endogenous and exogenous agents. The Streptomyces antibioticus glycosyltransferases, OleI and OleD, glycosylate and inactivate ole- andomycin and diverse macrolides including erythromycin, respec- tively. The structure of these enzyme–ligand complexes, in tandem with kinetic analysis of site-directed variants, provide insight into the interaction of macrolides with their synthetic apparatus. Erythromy- cin binds to OleD and the 23S RNA of its target ribosome in the same conformation and, although the antibiotic contains a large number of polar groups, its interaction with these macromolecules is primarily through hydrophobic contacts. Erythromycin and oleandomycin, when bound to OleD and OleI, respectively, adopt different confor- mations, reflecting a subtle effect on sugar positioning by virtue of a single change in the macrolide backbone. The data reported here provide structural insight into the mechanism of resistance to both endogenous and exogenous antibiotics, and will provide a platform for the future redesign of these catalysts for antibiotic remodelling. enzymology ͉ glycobiology ͉ erythromycin ͉ streptomyces ͉ glycoside icroorganisms, primarily Streptomyces, produce an array of Mclinically significant bioactive molecules that include antibi- Fig. 1. The mechanism of macrolide antibiotic resistance in S. antibioticus. otics and antitumor and immunosuppressant agents. Polyketides represent the most extensive group of bioactive molecules, which include macrolide polyketide antibiotics that target Gram-positive ensuring that high levels of the active form of the antimicrobial bacteria. Because resistance to these antimicrobial agents is not yet agent do not accumulate in the bacterium. The extracellular widespread among pathogens; they have been viewed as the ‘‘last ␤-glucosidase that activates oleandomycin is encoded by oleR, line of defense against bacterial pathogens’’ (World Health Orga- which is linked to oleI and the biosynthetic operon of the nization, January 30, 1998) (1). Macrolide antibiotics comprise a antibiotic (7). Unlike OleI, which only glycosylates oleandomy- macrocyclic backbone which following synthesis is decorated with cin, OleD displays broad acceptor specificity and hence will diverse glycans that play a key role in the specificity and potency of inactivate a wider range of macrolide antibiotics including tylosin these agents (2). and erythromycin, Fig. 2 (7, 10). Such promiscuity suggests that Streptomyces species that produce macrolide polyketide anti- OleD may function as a more general resistance mechanism, a biotics also use glycosylation to inactivate these molecules and view supported by the observation that oleD is not linked to the hence protect themselves from their own antimicrobial agents. In oleandomycin biosynthetic operon (12). Both OleI and OleD can this macrolide glycosyltransferase self-resistance mechanism, use several dinucleotide sugars as the donor substrate, although, intracellular glycosylation inactivates the antimicrobial agent, significantly, whereas OleI can mediate glycosyl transfer from which is subsequently reactivated, after secretion, by an extra- both UDP-␣-D-glucose and UDP-␣-D-galactose, OleD can use cellular ␤-glycosidase (Fig. 1) (3). The glycosylation of macro- lides is mediated by glycosyltransferases which transfer activated donor sugars to acceptor species (3–10). These enzymes are Author contributions: D.N.B., S.R., and M.R.P. contributed equally to this work; D.B., M.R.P., Ϸ G.J.D., and H.J.G. designed research; D.B., S.R., and M.R.P. performed research; D.B., M.R.P., grouped into 90 sequence-based families, and to date they J.P.T., E.J.D., C.M.-F., M.Y., and H.J.G. analyzed data; and B.G.D., G.J.D., and H.J.G. wrote the display only two major folds defined as GT-A and GT-B (see paper. below; http://afmb.cnrs-mrs.fr/CAZY/). In Streptomyces antibi- The authors declare no conflict of interest. oticus, the endogenous antibiotic oleandomycin is inactivated by This article is a PNAS Direct Submission. J.C. is a guest editor invited by the Editorial Board. two family 1 (GT-1; defined in ref. 11) glycosyltransferases, OleI Data deposition: The atomic coordinates have been deposited in the Protein Data Bank, and OleD, which share 45% sequence identity and catalyze www.pdb.org (PDB ID codes 2IYA and 2IYF). glucosyl transfer with inversion of anomeric configuration from §To whom correspondence may be addressed. E-mail: [email protected] or UDP-Glc to the O2 of the desosamine sugar of the antibiotic, [email protected]. Fig. 2 (9). OleI, which is tightly linked both genetically and This article contains supporting information online at www.pnas.org/cgi/content/full/ biochemically to the synthesis of the macrolide antibiotic ole- 0607897104/DC1. andomycin (7), displays a very low KM for the antibiotic, thus © 2007 by The National Academy of Sciences of the USA 5336–5341 ͉ PNAS ͉ March 27, 2007 ͉ vol. 104 ͉ no. 13 www.pnas.org͞cgi͞doi͞10.1073͞pnas.0607897104 Fig. 2. The reactions catalyzed by OleI and OleD. The enzymes transfer glucose from UDP-glucose to the O2 position of the desosamine sugar ap- pended to the macrolide backbone of macrolide antibiotics. The reaction occurs with inversion of anomeric configuration to generate the ␤-glucoside. Fig. 3. The three-dimensional structures of OleI (A) and OleD (B) shown as divergent (‘‘wall-eyed’’) stereo cartoons.
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