Structure to function of an α-glucan metabolic pathway that promotes Listeria monocytogenes pathogenesis

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Abstract

Here we employ a ‘systems structural biology’ approach to functionally characterize an unconventional α-glucan metabolic pathway from the food-borne pathogen Listeria monocytogenes (Lm). Crystal structure determination coupled with basic biochemical and biophysical assays allowed for the identification of anabolic, transport, catabolic and regulatory portions of the cycloalternan pathway. These findings provide numerous insights into cycloalternan pathway function and reveal the mechanism of repressor, open reading frame, kinase (ROK) transcription regulators. Moreover, by developing a structural overview we were able to anticipate the cycloalternan pathway's role in the metabolism of partially hydrolysed starch derivatives and demonstrate its involvement in Lm pathogenesis. These findings suggest that the cycloalternan pathway plays a role in interspecies resource competition—potentially within the host gastrointestinal tract—and establish the methodological framework for characterizing bacterial systems of unknown function.

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Figure 1: lmo2446lmo2444 operon function in extracellular CA anabolism.
Figure 2: lmo0178-lmo0184 operon function in cycloalternan (CA) uptake and intracellular catabolism.
Figure 3: Lmo0178 exerts transcriptional control over the lmo0178-lmo0184 operon.
Figure 4: The CA pathway promotes Lm infection and growth on partially hydrolysed starch derivatives.
Figure 5: Atomic model of CA pathway function.

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  • 14 July 2017

    In the PDF version of this article previously published, the year of publication provided in the footer of each page and in the 'How to cite' section was erroneously given as 2017, it should have been 2016. This error has now been corrected. The HTML version of the article was not affected.

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Acknowledgements

The Center for Structural Genomics of Infectious Diseases has been funded with federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health (NIH), Department of Health and Human Services, under contract nos. HHSN272200700058C and HHSN272201200026C (to W.F.A.). This work was supported by NIH grants R01 AI083241 and AI083241-03S1 (to N.E.F.) and F32 AI 115954 (to L.A.C.). Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under contract no. DE-AC02-06CH11357. Use of the LS-CAT Sector 21 was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor for the support of this research meprogram (grant no. 085P1000817). This work was supported by the Northwestern University High Throughput Analysis Laboratory, the Northwestern University Keck Biophysics Facility and a Cancer Center Support Grant (NCI CA060553). The authors thank C.-H. Luan, G. Minasov, Z. Wawrzak and B. Xayarath for assisting with experiments and S. Almo and G. Côté for providing reagents.

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S.H.L., L.A.C., N.E.F. and W.F.A. designed experiments. S.H.L., L.A.C. and A.S.H. performed experiments. S.H.L. wrote the manuscript.

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Correspondence to Wayne F. Anderson.

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Supplementary Figures 1–10, Supplementary Tables 1–3 and Supplementary References (PDF 1402 kb)

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Light, S., Cahoon, L., Halavaty, A. et al. Structure to function of an α-glucan metabolic pathway that promotes Listeria monocytogenes pathogenesis. Nat Microbiol 2, 16202 (2017). https://doi.org/10.1038/nmicrobiol.2016.202

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