Structure and mechanism of biosynthesis of Streptococcus mutans cell wall polysaccharide

Rush, Jeffrey S.; Zamakhaeva, Svetlana; Murner, Nicholas R.; Deng, Pan; Morris, Andrew J.; Kenner, Cameron W.; Black, Ian; Heiss, Christian; Azadi, Parastoo; Korotkov, Konstantin V.; Widmalm, Göran; Korotkova, Natalia

doi:10.1038/s41467-025-56205-1

Title: Structure and mechanism of biosynthesis of Streptococcus mutans cell wall polysaccharide

Journal Article · 21 January 2025 · Nature Communications

DOI: https://doi.org/10.1038/s41467-025-56205-1 · OSTI ID:2530521

Rush, Jeffrey S. ^[1];

^[1]; Murner, Nicholas R. ^[1]; Deng, Pan ^[1];

^[2]; Kenner, Cameron W. ^[1]; Black, Ian ^[3]; Heiss, Christian ^[3]; Azadi, Parastoo ^[3];

^[1];

^[4];

^[1]

University of Kentucky, Lexington, KY (United States)
University of Arkansas for Medical Science and Central Arkansas Veterans Affairs Healthcare System, Little Rock, AR (United States)
University of Georgia, Athens, GA (United States)
Stockholm University (Sweden)

Streptococcus mutans, the causative agent of human dental caries, expresses a cell wall attached Serotype c-specific Carbohydrate (SCC) that is critical for cell viability. SCC consists of a polyrhamnose backbone of →3)α-Rha(1 → 2)α-Rha(1→ repeats with glucose (Glc) side-chains and glycerol phosphate (GroP) decorations. This study reveals that SCC has one predominant and two more minor Glc modifications. The predominant Glc modification, α-Glc, attached to position 2 of 3-rhamnose, is installed by SccN and SccM glycosyltransferases and is the site of the GroP addition. The minor Glc modifications are β-Glc linked to position 4 of 3-rhamnose installed by SccP and SccQ glycosyltransferases, and α-Glc attached to position 4 of 2-rhamnose installed by SccN working in tandem with an unknown enzyme. Both the major and the minor β-Glc modifications control bacterial morphology, but only the GroP and major Glc modifications are critical for biofilm formation.

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Research Organization:: University of Georgia, Athens, GA (United States)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE; USDOE Office of Science (SC), Basic Energy Sciences (BES). Chemical Sciences, Geosciences & Biosciences Division (CSGB)

Grant/Contract Number:: SC0015662

OSTI ID:: 2530521

Journal Information:: Nature Communications, Journal Name: Nature Communications Journal Issue: 1 Vol. 16; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

References (57)

1H and13C NMR Data for 3-O-, 4-O- and 3,4-Di- 1O-glycosylated Methyl α-L-Rhamnopyranosides Khatuntseva, Elena A.; Shashkov, Alexander S.; Nifant’ev, Nikolay E. Magnetic Resonance in Chemistry, Vol. 35, Issue 6 https://doi.org/10.1002/(SICI)1097-458X(199706)35:6<414::AID-OMR74>3.0.CO;2-G	journal	June 1997
Wall Teichoic Acid Function, Biosynthesis, and Inhibition Swoboda, Jonathan G.; Campbell, Jennifer; Meredith, Timothy C. ChemBioChem, Vol. 11, Issue 1 https://doi.org/10.1002/cbic.200900557	journal	November 2009
Linkage Analysis of Oligosaccharides and Polysaccharides: A Tutorial Black, Ian; Heiss, Christian; Carlson, Russell W. Methods in Molecular Biology https://doi.org/10.1007/978-1-0716-1241-5_18	book	January 2021
The virulence of Streptococcus mutans and the ability to form biofilms Krzyściak, W.; Jurczak, A.; Kościelniak, D. European Journal of Clinical Microbiology & Infectious Diseases, Vol. 33, Issue 4 https://doi.org/10.1007/s10096-013-1993-7	journal	October 2013
Investigating the candidacy of the serotype specific rhamnan polysaccharide based glycoconjugates to prevent disease caused by the dental pathogen Streptococcus mutans St. Michael, Frank; Yang, Qingling; Cairns, Chantelle Glycoconjugate Journal, Vol. 35, Issue 1 https://doi.org/10.1007/s10719-017-9798-z	journal	October 2017
Synthesis, NMR, and conformational studies of methyl α-d-mannopyranoside 2-, 3-, 4-, and 6-monophosphates Bernlind, Christian; Oscarson, Stefan; Widmalm, Göran Carbohydrate Research, Vol. 263, Issue 2 https://doi.org/10.1016/0008-6215(94)00169-3	journal	October 1994
Polyprenyl phosphates: synthesis and structure-activity relationship for a biosynthetic system of Salmonella anatum O-specific polysaccharide Danilov, L. L.; Druzhinina, T. N.; Kalinchuk, N. A. Chemistry and Physics of Lipids, Vol. 51, Issue 3-4 https://doi.org/10.1016/0009-3084(89)90006-6	journal	November 1989
Characterization of the serotype e polysaccharide antigen of Streptococcus mutans Pritchard, David G.; Gregory, Richard L.; Michalek, Suzanne M. Molecular Immunology, Vol. 23, Issue 2 https://doi.org/10.1016/0161-5890(86)90035-0	journal	February 1986
Determination of the anomeric configuration of glycosyl esters of nucleoside pyrophosphates and polyisoprenyl phosphates by fast-atom bombardment tandem mass spectrometry Wolucka, Beata A.; de Hoffmann, Edmond; Rush, Jeffrey S. Journal of the American Society for Mass Spectrometry, Vol. 7, Issue 6 https://doi.org/10.1016/1044-0305(96)00020-7	journal	June 1996
Characterization of the group A streptococcal polysaccharide by two-dimensional 1H-nuclear-magnetic-resonance spectroscopy Huang, Dee Hua; Rama Krishna, N.; Pritchard, David G. Carbohydrate Research, Vol. 155 https://doi.org/10.1016/S0008-6215(00)90145-9	journal	November 1986
A Simple Method for the Isolation and Purification of Total Lipides from Animal Tissues Folch, Jordi; Lees, M.; Stanley, G. H. Sloane Journal of Biological Chemistry, Vol. 226, Issue 1 https://doi.org/10.1016/S0021-9258(18)64849-5	journal	May 1957
Isolation and analysis of cell wall components from Streptococcus pneumoniae Bui, Nhat Khai; Eberhardt, Alice; Vollmer, Daniela Analytical Biochemistry, Vol. 421, Issue 2 https://doi.org/10.1016/j.ab.2011.11.026	journal	February 2012
A malachite green-based assay to assess glucan phosphatase activity Sherwood, Amanda R.; Paasch, Bradley C.; Worby, Carolyn A. Analytical Biochemistry, Vol. 435, Issue 1 https://doi.org/10.1016/j.ab.2012.10.044	journal	April 2013
Novel inhibitors of Mycobacterium tuberculosis dTDP-6-deoxy-l-lyxo-4-hexulose reductase (RmlD) identified by virtual screening Wang, Yi; Hess, Tamara Noelle; Jones, Victoria Bioorganic & Medicinal Chemistry Letters, Vol. 21, Issue 23 https://doi.org/10.1016/j.bmcl.2011.09.094	journal	December 2011
Elucidating the role of N-acetylglucosamine in Group A Carbohydrate for the development of an effective glycoconjugate vaccine against Group A Streptococcus Pitirollo, Olimpia; Di Benedetto, Roberta; Henriques, Pedro Carbohydrate Polymers, Vol. 311 https://doi.org/10.1016/j.carbpol.2023.120736	journal	July 2023
NMR chemical shift prediction and structural elucidation of linker-containing oligo- and polysaccharides using the computer program CASPER Dorst, Kevin M.; Widmalm, Göran Carbohydrate Research, Vol. 533 https://doi.org/10.1016/j.carres.2023.108937	journal	November 2023
The Classical Lancefield Antigen of Group A Streptococcus Is a Virulence Determinant with Implications for Vaccine Design van Sorge, Nina M.; Cole, Jason N.; Kuipers, Kirsten Cell Host & Microbe, Vol. 15, Issue 6 https://doi.org/10.1016/j.chom.2014.05.009	journal	June 2014
Quantitative high-performance liquid chromatography analysis of the pool levels of undecaprenyl phosphate and its derivatives in bacterial membranes Barreteau, Hélène; Magnet, Sophie; Ghachi, Meriem El Journal of Chromatography B, Vol. 877, Issue 3 https://doi.org/10.1016/j.jchromb.2008.12.010	journal	January 2009
Modifications of cell wall polymers in Gram-positive bacteria by multi-component transmembrane glycosylation systems Rismondo, Jeanine; Gillis, Annika; Gründling, Angelika Current Opinion in Microbiology, Vol. 60 https://doi.org/10.1016/j.mib.2021.01.007	journal	April 2021
Evaluation of a Group A Streptococcus synthetic oligosaccharide as vaccine candidate Kabanova, Anna; Margarit, Immaculada; Berti, Francesco Vaccine, Vol. 29, Issue 1 https://doi.org/10.1016/j.vaccine.2010.09.018	journal	December 2010
Primary Structure of Glycans by NMR Spectroscopy Fontana, Carolina; Widmalm, Göran Chemical Reviews, Vol. 123, Issue 3 https://doi.org/10.1021/acs.chemrev.2c00580	journal	January 2023
Immunogenicity Assessment of Cell Wall Carbohydrates of Group A Streptococcus via Self-Adjuvanted Glyco-lipopeptides Khatun, Farjana; Dai, Charles C.; Rivera-Hernandez, Tania ACS Infectious Diseases, Vol. 7, Issue 2 https://doi.org/10.1021/acsinfecdis.0c00722	journal	February 2021
Group A Streptococcus Cell Wall Oligosaccharide-Streptococcal C5a Peptidase Conjugates as Effective Antibacterial Vaccines Wang, Subo; Zhao, Yisheng; Wang, Guirong ACS Infectious Diseases, Vol. 6, Issue 2 https://doi.org/10.1021/acsinfecdis.9b00347	journal	December 2019
Amphomycin: effect of the lipopeptide antibiotic on the glycosylation and extraction of dolichyl monophosphate in calf brain membranes Banerjee, Dipak K.; Scher, Malka G.; Waechter, C. J. Biochemistry, Vol. 20, Issue 6 https://doi.org/10.1021/bi00509a024	journal	March 1981
Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants the Keio collection Baba, Tomoya; Ara, Takeshi; Hasegawa, Miki Molecular Systems Biology, Vol. 2, Article No. 2006.0008 https://doi.org/10.1038/msb4100050	journal	February 2006
PplD is a de-N-acetylase of the cell wall linkage unit of streptococcal rhamnopolysaccharides Rush, Jeffrey S.; Parajuli, Prakash; Ruda, Alessandro Nature Communications, Vol. 13, Issue 1 https://doi.org/10.1038/s41467-022-28257-0	journal	February 2022
Discovery of glycerol phosphate modification on streptococcal rhamnose polysaccharides Edgar, Rebecca J.; van Hensbergen, Vincent P.; Ruda, Alessandro Nature Chemical Biology, Vol. 15, Issue 5 https://doi.org/10.1038/s41589-019-0251-4	journal	April 2019
Modification of cell wall polysaccharide guides cell division in Streptococcus mutans Zamakhaeva, Svetlana; Chaton, Catherine T.; Rush, Jeffrey S. Nature Chemical Biology, Vol. 17, Issue 8 https://doi.org/10.1038/s41589-021-00803-9	journal	May 2021
Glycosyltransferases: mechanisms and applications in natural product development Liang, Dong-Mei; Liu, Jia-Heng; Wu, Hao Chemical Society Reviews, Vol. 44, Issue 22 https://doi.org/10.1039/C5CS00600G	journal	January 2015
Molecular basis for recognition of the Group A Carbohydrate backbone by the PlyC streptococcal bacteriophage endolysin King, Harley; Ajay Castro, Sowmya; Pohane, Amol Arunrao Biochemical Journal, Vol. 478, Issue 12 https://doi.org/10.1042/BCJ20210158	journal	June 2021
The molecular mechanism of N -acetylglucosamine side-chain attachment to the Lancefield group A carbohydrate in Streptococcus pyogenes Rush, Jeffrey S.; Edgar, Rebecca J.; Deng, Pan Journal of Biological Chemistry, Vol. 292, Issue 47 https://doi.org/10.1074/jbc.M117.815910	journal	October 2017
Group A, B, C, and G Streptococcus Lancefield antigen biosynthesis is initiated by a conserved α-d-GlcNAc-β-1,4-l-rhamnosyltransferase Zorzoli, Azul; Meyer, Benjamin H.; Adair, Elaine Journal of Biological Chemistry, Vol. 294, Issue 42 https://doi.org/10.1074/jbc.RA119.009894	journal	October 2019
Regulatory mechanisms of exopolysaccharide synthesis and biofilm formation in Streptococcus mutans Zheng, Ting; Jing, Meiling; Gong, Tao Journal of Oral Microbiology, Vol. 15, Issue 1 https://doi.org/10.1080/20002297.2023.2225257	journal	June 2023
Group A Streptococcus (GAS) Carbohydrate as an Immunogen for Protection against GAS Infection Sabharwal, Hemant; Michon, Francis; Nelson, Daniel The Journal of Infectious Diseases, Vol. 193, Issue 1 https://doi.org/10.1086/498618	journal	January 2006
Bacterial glycobiology: rhamnose-containing cell wall polysaccharides in Gram-positive bacteria Mistou, Michel-Yves; Sutcliffe, Iain C.; van Sorge, Nina M. FEMS Microbiology Reviews, Vol. 40, Issue 4 https://doi.org/10.1093/femsre/fuw006	journal	March 2016
Rapid degradation of Streptococcus pyogenes biofilms by PlyC, a bacteriophage-encoded endolysin Shen, Yang; Köller, Thomas; Kreikemeyer, Bernd Journal of Antimicrobial Chemotherapy, Vol. 68, Issue 8 https://doi.org/10.1093/jac/dkt104	journal	April 2013
Streptococcal dTDP‐L‐rhamnose biosynthesis enzymes: functional characterization and lead compound identification van der Beek, Samantha L.; Zorzoli, Azul; Çanak, Ebru Molecular Microbiology, Vol. 111, Issue 4 https://doi.org/10.1111/mmi.14197	journal	January 2019
Streptococcus mutansrequires mature rhamnose‐glucose polysaccharides for proper pathophysiology, morphogenesis and cellular division Kovacs, Christopher J.; Faustoferri, Roberta C.; Bischer, Andrew P. Molecular Microbiology, Vol. 112, Issue 3 https://doi.org/10.1111/mmi.14330	journal	July 2019
High avidity drives the interaction between the streptococcal C1 phage endolysin, PlyC, with the cell surface carbohydrates of Group A Streptococcus Broendum, Sebastian S.; Williams, Daniel E.; Hayes, Brooke K. Molecular Microbiology, Vol. 116, Issue 2 https://doi.org/10.1111/mmi.14719	journal	April 2021
Exploiting the Reversibility of Natural Product Glycosyltransferase-Catalyzed Reactions Zhang, C. Science, Vol. 313, Issue 5791 https://doi.org/10.1126/science.1130028	journal	September 2006
Deficiency of RgpG Causes Major Defects in Cell Division and Biofilm Formation, and Deficiency of LytR-CpsA-Psr Family Proteins Leads to Accumulation of Cell Wall Antigens in Culture Medium by Streptococcus mutans De, Arpan; Liao, Sumei; Bitoun, Jacob P. Applied and Environmental Microbiology, Vol. 83, Issue 17 https://doi.org/10.1128/AEM.00928-17	journal	July 2017
Glycosyltransferase-Mediated Biofilm Matrix Dynamics and Virulence of Streptococcus mutans Rainey, Katherine; Michalek, Suzanne M.; Wen, Zezhang T. Applied and Environmental Microbiology, Vol. 85, Issue 5 https://doi.org/10.1128/AEM.02247-18	journal	March 2019
Expression and Characterization of Streptococcal rgp Genes Required for Rhamnan Synthesis in Escherichia coli Shibata, Yukie; Yamashita, Yoshihisa; Ozaki, Kazuhisa Infection and Immunity, Vol. 70, Issue 6 https://doi.org/10.1128/IAI.70.6.2891-2898.2002	journal	June 2002
A Defective Undecaprenyl Pyrophosphate Synthase Induces Growth and Morphological Defects That Are Suppressed by Mutations in the Isoprenoid Pathway of Escherichia coli MacCain, William J.; Kannan, Suresh; Jameel, Dannah Z. Journal of Bacteriology, Vol. 200, Issue 18 https://doi.org/10.1128/JB.00255-18	journal	September 2018
RgpF Is Required for Maintenance of Stress Tolerance and Virulence in Streptococcus mutans Kovacs, C. J.; Faustoferri, R. C.; Quivey, R. G. Journal of Bacteriology, Vol. 199, Issue 24 https://doi.org/10.1128/JB.00497-17	journal	December 2017
Depletion of Undecaprenyl Pyrophosphate Phosphatases Disrupts Cell Envelope Biogenesis in Bacillus subtilis Zhao, Heng; Sun, Yingjie; Peters, Jason M. Journal of Bacteriology, Vol. 198, Issue 21 https://doi.org/10.1128/JB.00507-16	journal	August 2016
Disruption of l -Rhamnose Biosynthesis Results in Severe Growth Defects in Streptococcus mutans Bischer, Andrew P.; Kovacs, Christopher J.; Faustoferri, Roberta C. Journal of Bacteriology, Vol. 202, Issue 6 https://doi.org/10.1128/JB.00728-19	journal	February 2020
A Novel Gene Required for Rhamnose-Glucose Polysaccharide Synthesis in Streptococcus mutans Yamashita, Yoshihisa; Shibata, Yukie; Nakano, Yoshio Journal of Bacteriology, Vol. 181, Issue 20 https://doi.org/10.1128/JB.181.20.6556-6559.1999	journal	October 1999
Analysis of Loci Required for Determination of Serotype Antigenicity in Streptococcus mutans and Its Clinical Utilization Shibata, Yukie; Ozaki, Kazuhisa; Seki, Mitsuko Journal of Clinical Microbiology, Vol. 41, Issue 9 https://doi.org/10.1128/JCM.41.9.4107-4112.2003	journal	September 2003
Demonstration of Streptococcus mutans with a Cell Wall Polysaccharide Specific to a New Serotype, k , in the Human Oral Cavity Nakano, Kazuhiko; Nomura, Ryota; Nakagawa, Ichiro Journal of Clinical Microbiology, Vol. 42, Issue 1 https://doi.org/10.1128/JCM.42.1.198-202.2004	journal	January 2004
The Biology of Streptococcus mutans Lemos, J. A.; Palmer, S. R.; Zeng, L. Microbiology Spectrum, Vol. 7, Issue 1 https://doi.org/10.1128/microbiolspec.gpp3-0051-2018	journal	February 2019
Role of Streptococcus mutans in human dental decay Loesche, W. J. Microbiological Reviews, Vol. 50, Issue 4 https://doi.org/10.1128/mr.50.4.353-380.1986	journal	December 1986
A Rapid Method of Total Lipid Extraction and Purification Bligh, E. G.; Dyer, W. J. Canadian Journal of Biochemistry and Physiology, Vol. 37, Issue 1 https://doi.org/10.1139/y59-099	journal	January 1959
Role of the Group B Antigen of Streptococcus agalactiae: A Peptidoglycan-Anchored Polysaccharide Involved in Cell Wall Biogenesis Caliot, Élise; Dramsi, Shaynoor; Chapot-Chartier, Marie-Pierre PLoS Pathogens, Vol. 8, Issue 6 https://doi.org/10.1371/journal.ppat.1002756	journal	June 2012
Serotype classification of Streptococcus mutans and its detection outside the oral cavity Nakano, Kazuhiko; Ooshima, Takashi Future Microbiology, Vol. 4, Issue 7 https://doi.org/10.2217/fmb.09.64	journal	September 2009
Rational Design of a Glycoconjugate Vaccine against Group A Streptococcus Di Benedetto, Roberta; Mancini, Francesca; Carducci, Martina International Journal of Molecular Sciences, Vol. 21, Issue 22 https://doi.org/10.3390/ijms21228558	journal	November 2020
A bacteriophage endolysin that eliminates intracellular streptococci Shen, Yang; Barros, Marilia; Vennemann, Tarek eLife, Vol. 5 https://doi.org/10.7554/eLife.13152	journal	March 2016

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Title: Structure and mechanism of biosynthesis of Streptococcus mutans cell wall polysaccharide

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