Identification, function and structure of the mycobacterial sulfotransferase that initiates sulfolipid-1 biosynthesis

Mougous, Joseph D.; Kim, Young-Mo; Senaratne, Ryan H.; Lee, Dong H; Akey, D.L.; Lin, Fiona L.; Munchel, Sarah E.; Pratt, Matthew R.; Riley, Lee W.; Leary, Julie A.; Berger, James M.; Bertozzi, Carolyn R.

doi:10.1038/nsmb802

Cited by 100 publications

(95 citation statements)

References 68 publications

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“…⌬cysC was successfully complemented (⌬cysC-ctrl) by expressing only the APS kinase domain of CysNC. To confirm that the defect in SL-1 biosynthesis was due to a lack of sulfation, we also analyzed extracts from ⌬cysC and WT for the first committed sulfated precursor of SL-1, trehalose-2-sulfate (24). As expected, ⌬cysC did not contain detectable levels of trehalose-2-sulfate (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To validate the mutant, we analyzed extracts for the presence of sulfated SL-1, the biosynthesis of which has been shown to rely on a PAPS-dependent sulfotransferase (24). Extracts from ⌬cysC were devoid of SL-1 as determined by both Fourier transform ion cyclotron resonance (FT-ICR) MS (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…SL-1 has generated much interest because of its elaborate structure and the observation that its abundance correlates with strain virulence (15)(16)(17)(18)(19)(20)(21)(22). Advances in M. tuberculosis genetics and genome sequence data facilitated several contemporary studies that addressed aspects of the biosynthesis and the function of SL-1 in vivo (23)(24)(25)(26). Although these studies have greatly increased our understanding of SL-1, its function in the life cycle of the bacterium remains unknown.…”

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confidence: 99%

“…Because the sulfate ester is generally a determinant of function, these enzymes are attractive targets for the perturbation and study of sulfated molecules. In previous reports, we used conserved sulfotransferase motifs and mycobacterial genome sequence information to identify four sulfotransferases (stf0-3) in M. tuberculosis (11,24). The abundance of stf genes suggested that M. tuberculosis has the capacity to synthesize sulfated metabolites in addition to SL-1.…”

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confidence: 99%

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A sulfated metabolite produced bystf3negatively regulates the virulence ofMycobacteriumtuberculosis

Mougous

Senaratne

Kim

et al. 2006

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

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Sulfated molecules have been shown to modulate isotypic interactions between cells of metazoans and heterotypic interactions between bacterial pathogens or symbionts and their eukaryotic host cells. Mycobacterium tuberculosis, the causative agent of tuberculosis, produces sulfated molecules that have eluded functional characterization for decades. We demonstrate here that a previously uncharacterized sulfated molecule, termed S881, is localized to the outer envelope of M. tuberculosis and negatively regulates the virulence of the organism in two mouse infection models. Furthermore, we show that the biosynthesis of S881 relies on the universal sulfate donor 3 -phosphoadenosine-5 -phosphosulfate and a previously uncharacterized sulfotransferase, stf3. These findings extend the known functions of sulfated molecules as general modulators of cell-cell interactions to include those between a bacterium and a human host.Fourier transform ion cyclotron resonance ͉ hypervirulent ͉ sulfate assimilation ͉ kinase ͉ adenosine-5-phosphosulfate A wide variety of organisms use sulfated molecules to control extracellular events. In mammals, sulfation of tyrosine residues on cell surface proteins is important for the interactions of chemokines with certain chemokine receptors, and for viral binding and entry (1-6). Sulfated glycans modulate processes such as leukocyte homing to lymph nodes, clearance of serum glycoproteins, and blood coagulation (7-9). Members of the glypican family that are modified with sulfated glycosaminoglycans guide organ development in Drosophila by maintaining a morphogen concentration gradient (10).In bacteria, sulfated glycolipids have been shown to serve as extracellular signaling molecules (11). The nitrogen fixing bacterium Sinorhizobium meliloti secretes the nodulation factor NodRm-1, a tetrasaccharide bearing both sulfate and lipid modifications (12). This molecule binds a receptor on the host plant, normally alfalfa, and induces root nodule formation. The sulfate group is critical for the function of NodRm-1, because the unsulfated form fails to induce root nodulation in alfalfa. In the rice blight-causing pathogen Xanthomonas oryzae, several genes involved in the synthesis of sulfated metabolites have been identified as avirulence factors with respect to certain host strains (13,14). These examples suggest that bacterial sulfated metabolites can participate in dialogue with eukaryotic hosts, analogous to their role in mammalian cell-cell communication.Mycobacteria produce an unusually complex array of sulfated structures (11). Sulfolipid-1 (SL-1), an abundant component of the cell envelope of M. tuberculosis, is the best characterized of these molecules. SL-1 has generated much interest because of its elaborate structure and the observation that its abundance correlates with strain virulence (15)(16)(17)(18)(19)(20)(21)(22). Advances in M. tuberculosis genetics and genome sequence data facilitated several contemporary studies that addressed aspects of the biosynthesis and the function of SL-1 in v...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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A sulfated metabolite produced bystf3negatively regulates the virulence ofMycobacteriumtuberculosis

Mougous

Senaratne

Kim

et al. 2006

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

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“…[8][9][10][11][12] By contrast, significant progress has been made in the last 5 years in understanding the biosynthesis of SL-I. [13][14][15][16][17][18][19][20][21] One of its presumed precursors appears to have a hydroxyphthioceranoyl group at the 3′ position (rather than the 6′ position as shown in Figure 1), however, casting some doubt on the accuracy of Goren's structure. 19,20 To investigate the biological activity of SL-I and to resolve questions concerning its precise structure, we set out to synthesize SL-I and analogs thereof.…”

Section: Introductionmentioning

confidence: 99%

Synthetic Studies toward Mycobacterium tuberculosis Sulfolipid-I

Leigh

Bertozzi

2008

J. Org. Chem.

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Sulfolipid-I (SL-I) is an abundant metabolite found in the cell wall of Mycobacterium tuberculosis that is comprised of a trehalose 2-sulfate core modified with four fatty acyl substituents. The correlation of its abundance with the virulence of clinical isolates suggests a role for SL-I in pathogenesis, although its biological functions remain unknown. Here we describe the synthesis of a SL-I analogue bearing unnatural lipid substituents. A key feature of the synthesis was application of an intramolecular aglycon delivery reaction to join two differentially protected glucose monomers, one prepared with a novel α-selective glycosylation. The route developed for the model compound can be readily extended to the synthesis of native SL-I as well as additional analogues for use in the investigation of SL-I's functions.

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