Hierarchical Glycolytic Pathways Control the Carbohydrate Utilization Regulator in Human Gut

Overview

Journal bioRxiv

Date 2024 Nov 28

PMID 39605394

Authors

Seth Kabonick

Kamalesh Verma

Jennifer L Modesto

Victoria H Pearce

Kailyn M Winokur

Eduardo A Groisman

Guy E Townsend

Affiliations

Soon will be listed here.

Abstract

Human dietary choices control the gut microbiome. Industrialized populations consume abundant glucose and fructose, resulting in microbe-dependent intestinal disorders. Simple sugars inhibit the carbohydrate utilization regulator (Cur), a transcription factor in the prominent gut bacterial phylum, . Cur encodes products necessary for carbohydrate utilization, host immunomodulation, and intestinal colonization. Here, we demonstrate how simple sugars decrease Cur activity in the mammalian gut. Our findings in two species show that ATP-dependent fructose-1,6-bisphosphate (FBP) synthesis is necessary for glucose or fructose to inhibit Cur, but dispensable for growth because of an essential pyrophosphate (PPi)-dependent enzyme. Furthermore, we show that ATP-dependent FBP synthesis is required to regulate Cur in the gut but does not contribute to fitness when is absent, indicating PPi is sufficient to drive glycolysis in these bacteria. Our findings reveal how sugar-rich diets inhibit Cur, thereby disrupting fitness and diminishing products that are beneficial to the host.

References

Siegel L, Hylemon P, Phibbs Jr P . Cyclic adenosine 3',5'-monophosphate levels and activities of adenylate cyclase and cyclic adenosine 3',5'-monophosphate phosphodiesterase in Pseudomonas and Bacteroides. J Bacteriol. 1977; 129(1):87-96. PMC: 234899. DOI: 10.1128/jb.129.1.87-96.1977. View

Modesto J, Pearce V, Townsend 2nd G . Harnessing gut microbes for glycan detection and quantification. Nat Commun. 2023; 14(1):275. PMC: 9845299. DOI: 10.1038/s41467-022-35626-2. View

Stulke J, Hillen W . Carbon catabolite repression in bacteria. Curr Opin Microbiol. 1999; 2(2):195-201. DOI: 10.1016/S1369-5274(99)80034-4. View

LOOMIS Jr W, MAGASANIK B . The catabolite repression gene of the lac operon in Escherichia coli. J Mol Biol. 1967; 23(3):487-94. DOI: 10.1016/s0022-2836(67)80120-7. View

Townsend 2nd G, Han W, Schwalm 3rd N, Hong X, Bencivenga-Barry N, Goodman A . A Master Regulator of Bacteroides thetaiotaomicron Gut Colonization Controls Carbohydrate Utilization and an Alternative Protein Synthesis Factor. mBio. 2020; 11(1). PMC: 6989115. DOI: 10.1128/mBio.03221-19. View

Simoni R, Roseman S, Saier Jr M . Sugar transport. Properties of mutant bacteria defective in proteins of the phosphoenolpyruvate: sugar phosphotransferase system. J Biol Chem. 1976; 251(21):6584-97. View

Barabote R, Saier Jr M . Comparative genomic analyses of the bacterial phosphotransferase system. Microbiol Mol Biol Rev. 2005; 69(4):608-34. PMC: 1306802. DOI: 10.1128/MMBR.69.4.608-634.2005. View

Garcia-Bayona L, Comstock L . Streamlined Genetic Manipulation of Diverse and Isolates from the Human Gut Microbiota. mBio. 2019; 10(4). PMC: 6692515. DOI: 10.1128/mBio.01762-19. View

Carreon-Rodriguez O, Gosset G, Escalante A, Bolivar F . Glucose Transport in : From Basics to Transport Engineering. Microorganisms. 2023; 11(6). PMC: 10305011. DOI: 10.3390/microorganisms11061588. View

10.

Saier Jr M, Feucht B, Hofstadter L . Regulation of carbohydrate uptake and adenylate cyclase activity mediated by the enzymes II of the phosphoenolpyruvate: sugar phosphotransferase system in Escherichia coli. J Biol Chem. 1976; 251(3):883-92. View

11.

Jang C, Hui S, Lu W, Cowan A, Morscher R, Lee G . The Small Intestine Converts Dietary Fructose into Glucose and Organic Acids. Cell Metab. 2018; 27(2):351-361.e3. PMC: 6032988. DOI: 10.1016/j.cmet.2017.12.016. View

12.

Bry L, Falk P, Midtvedt T, Gordon J . A model of host-microbial interactions in an open mammalian ecosystem. Science. 1996; 273(5280):1380-3. DOI: 10.1126/science.273.5280.1380. View

13.

Goodman A, McNulty N, Zhao Y, Leip D, Mitra R, Lozupone C . Identifying genetic determinants needed to establish a human gut symbiont in its habitat. Cell Host Microbe. 2009; 6(3):279-89. PMC: 2895552. DOI: 10.1016/j.chom.2009.08.003. View

14.

Koropatkin N, Martens E, Gordon J, Smith T . Starch catabolism by a prominent human gut symbiont is directed by the recognition of amylose helices. Structure. 2008; 16(7):1105-15. PMC: 2563962. DOI: 10.1016/j.str.2008.03.017. View

15.

Pearce V, Groisman E, Townsend 2nd G . Dietary sugars silence the master regulator of carbohydrate utilization in human gut Bacteroides species. Gut Microbes. 2023; 15(1):2221484. PMC: 10294740. DOI: 10.1080/19490976.2023.2221484. View

16.

Hylemon P, Phibbs Jr P . Evidence against the presence of cyclic AMP and related enzymes in selected strains of Bacteroides fragilis. Biochem Biophys Res Commun. 1974; 60(1):88-95. DOI: 10.1016/0006-291x(74)90176-4. View

17.

Bruckner R, Titgemeyer F . Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization. FEMS Microbiol Lett. 2002; 209(2):141-8. DOI: 10.1111/j.1574-6968.2002.tb11123.x. View

18.

Townsend 2nd G, Han W, Schwalm 3rd N, Raghavan V, Barry N, Goodman A . Dietary sugar silences a colonization factor in a mammalian gut symbiont. Proc Natl Acad Sci U S A. 2018; 116(1):233-238. PMC: 6320540. DOI: 10.1073/pnas.1813780115. View

19.

MAGASANIK B . Catabolite repression. Cold Spring Harb Symp Quant Biol. 1961; 26:249-56. DOI: 10.1101/sqb.1961.026.01.031. View

20.

Lovingshimer M, Siegele D, Reinhart G . Construction of an inducible, pfkA and pfkB deficient strain of Escherichia coli for the expression and purification of phosphofructokinase from bacterial sources. Protein Expr Purif. 2005; 46(2):475-82. DOI: 10.1016/j.pep.2005.09.015. View