Dissection of Hexosyl- and Sialyltransferase Domains in the Bifunctional Capsule Polymerases from Neisseria meningitidis W and Y Defines a New Sialyltransferase Family

Romanow, Angela; Keys, Timothy G.; Stummeyer, Katharina; Freiberger, Friedrich; Henrissat, Bernard; Gerardy‐Schahn, Rita

doi:10.1074/jbc.m114.597773

Cited by 22 publications

(30 citation statements)

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“…However, relative activity assays showed that the H265A and P266A mutants maintain reduced catalytic activity. Another bacterial "sialyl motif" S (S/T) (30,31) is reduced to a single invariant Ser residue (S300 in WbbB Rt ) in most β-Kdo GTs, although the four closest WbbB homologs also possess an adjacent Ser residue. Mutation S300A dramatically decreases enzyme activity, whereas S301A only has a slight catalytic defect.…”

Section: Discussionmentioning

confidence: 99%

Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)

Ovchinnikova

Mallette

Koizumi

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Kdo (3-deoxy-D-manno-oct-2-ulosonic acid) is an eight-carbon sugar mostly confined to Gram-negative bacteria. It is often involved in attaching surface polysaccharides to their lipid anchors. α-Kdo provides a bridge between lipid A and the core oligosaccharide in all bacterial LPSs, whereas an oligosaccharide of β-Kdo residues links "group 2" capsular polysaccharides to (lyso)phosphatidylglycerol. β-Kdo is also found in a small number of other bacterial polysaccharides. The structure and function of the prototypical cytidine monophosphate-Kdo-dependent α-Kdo glycosyltransferase from LPS assembly is well characterized. In contrast, the β-Kdo counterparts were not identified as glycosyltransferase enzymes by bioinformatics tools and were not represented among the 98 currently recognized glycosyltransferase families in the Carbohydrate-Active Enzymes database. We report the crystallographic structure and function of a prototype β-Kdo GT from WbbB, a modular protein participating in LPS O-antigen synthesis in Raoultella terrigena. The β-Kdo GT has dual Rossmann-fold motifs typical of GT-B enzymes, but extensive deletions, insertions, and rearrangements result in a unique architecture that makes it a prototype for a new GT family (GT99). The cytidine monophosphate-binding site in the C-terminal α/β domain closely resembles the corresponding site in bacterial sialyltransferases, suggesting an evolutionary connection that is not immediately evident from the overall fold or sequence similarities. microbial glycobiology | 3-deoxy-D-manno-oct-2-ulosonic acid | Kdo | glycosyltransferase | polysaccharide

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Section: Discussionmentioning

confidence: 99%

Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)

Ovchinnikova

Mallette

Koizumi

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…Other two-domain CPs consist of two equally folded domains, like the GT-A folded GAG CPS synthases isolated from P. multocida (58,59) and E. coli K4 (54) or the GT-B folded CPs from N. meningitidis serogroups W and Y (39). All of these earlier described enzymes assemble CPSs consisting of dimeric repeating units that only contain glycosidic linkages, and they use two different nucleotide sugars as donors.…”

Section: Discussionmentioning

confidence: 99%

“…It is most likely that this region, for which no homology could be found using PHYRE2, participates in the catalytic activity. A similar region was identified in the CPs of N. meningitidis serogroups W and Y and shown to contribute to the formation of the functionally active sialyltransferase domain (36,39). To precisely annotate functions to each domain in CslB, more studies are needed that require the availability of suited, preferably fluorescently labeled acceptors to enable the recording of single transfer reactions.…”

Section: Volume 290 • Number 40 • October 2 2015mentioning

confidence: 99%

“…In this context, we focused our work on capsule polymerases (CPs) of both pathogenic and rarely occurring meningococcal serogroups and thereby follow two research lines: (i) exploitation of recombinant CPs for in vitro production of glycoconjugate vaccines (30,31) and (ii) delineation of structure-function relationships as a basis for drug target evaluation (32)(33)(34)(35)(36)(37)(38)(39). Recently, we cloned and functionally expressed the CPs from serogroups A (CsaB) and X (CsxA) (30,31).…”

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confidence: 99%

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The Capsule Polymerase CslB of Neisseria meningitidis Serogroup L Catalyzes the Synthesis of a Complex Trimeric Repeating Unit Comprising Glycosidic and Phosphodiester Linkages

Litschko

Romano

Pinto

et al. 2015

Journal of Biological Chemistry

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Background:The trimeric repeating unit forming the capsular polysaccharide of Neisseria meningitidis serogroup L comprises glycosidic and phosphodiester bonds. Results: We identified the two-domain capsule polymerase CslB of N. meningitidis serogroup L to assemble this complex trimer with UDP-GlcNAc as substrate. Conclusion: CslB represents a unique capsule polymerase in group 2 capsule expressing bacteria. Significance: Understanding capsule biosynthesis is mandatory for combatting encapsulated bacterial pathogens.

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“…Csy and Csw, which catalyze heteropolymers, are 98% identical to each other at the nucleotide level, but they are only ~12% identical to Csc and Csb. Csw is a chimeric enzyme comprising two independent sialyl- and galactosyltransferase catalytic domains that work in succession because Csw variants with only one functionality can complement each other in trans when combined in vitro (Romanow et al, 2013; Romanow et al, 2014). Interestingly, amino acid residue 310, located within the predicted hexosyltransferase moiety of the polymerase, accounted for substrate specificity of W and Y capsules: Pro-310 determined galactosyltransferase activity that resulted in a serogroup W capsule, while Gly-310 determined glucosyltransferase activity that yielded a serogroup Y capsule (Claus et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Regulation of capsule inNeisseria meningitidis

Tzeng

Thomas

Stephens

2015

Critical Reviews in Microbiology

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Neisseria meningitidis , a devastating pathogen exclusive to humans, expresses capsular polysaccharides that are the major meningococcal virulence determinants and the basis for successful meningococcal vaccines. With rare exceptions, the expression of capsule (serogroups A, B, C, W, X, Y) is required for systemic invasive meningococcal disease. Changes in capsule expression or structure (e.g. hypo- or hyperencapsulation, capsule “switching”, acetylation) can influence immunologic diagnostic assays or lead to immune escape. The loss or down-regulation of capsule is also critical in meningococcal biology facilitating meningococcal attachment, microcolony formation and the carriage state at human mucosal surfaces. Encapsulated meningococci contain a cps locus with promoters located in an intergenic region between the biosynthesis and the conserved capsule transport operons. The cps intergenic region is transcriptionally regulated (and thus the amount of capsule expressed) by IS element insertion, by a two-component system, MisR/MisS, and through sequence changes that result in post transcriptional RNA thermoregulation. Reversible on-off phase variation of capsule expression is controlled by slipped strand mispairing of homo-polymeric tracts and by precise insertion and excision of IS elements (e.g. IS1301) in the biosynthesis operon. Capsule structure can be altered by phase-variable expression of capsular polymer modification enzymes or “switched” through transformation and homologous recombination of different polymerases. Understanding the complex regulation of meningococcal capsule has important implications for meningococcal biology, pathogenesis, diagnostics, current and future vaccine development and vaccine strategies.

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Dissection of Hexosyl- and Sialyltransferase Domains in the Bifunctional Capsule Polymerases from Neisseria meningitidis W and Y Defines a New Sialyltransferase Family

Cited by 22 publications

References 40 publications

Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)

Bacterial β-Kdo glycosyltransferases represent a new glycosyltransferase family (GT99)

The Capsule Polymerase CslB of Neisseria meningitidis Serogroup L Catalyzes the Synthesis of a Complex Trimeric Repeating Unit Comprising Glycosidic and Phosphodiester Linkages

Regulation of capsule inNeisseria meningitidis

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