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Co-culturing experiments reveal the uptake of myo-inositol phosphate synthase (EC 5.5.1.4) in an inositol auxotroph of Saccharomyces cerevisiae

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dc.contributor.authorSteele, Erika
dc.contributor.authorAlebous, Hana D.
dc.contributor.authorVickers, Macy
dc.contributor.authorHarris, Mary E.
dc.contributor.authorJohnson, Margaret D.
dc.contributor.otherUniversity of Alabama Tuscaloosa
dc.contributor.otherUniversity of Jordan
dc.date.accessioned2023-09-28T19:11:38Z
dc.date.available2023-09-28T19:11:38Z
dc.date.issued2021
dc.description.abstractBackgroundMyo-Inositol Phosphate Synthase (MIP) catalyzes the conversion of glucose 6- phosphate into inositol phosphate, an essential nutrient and cell signaling molecule. Data obtained, first in bovine brain and later in plants, established MIP expression in organelles and in extracellular environments. A physiological role for secreted MIP has remained elusive since its first detection in intercellular space. To provide further insight into the role of MIP in intercellular milieus, we tested the hypothesis that MIP may function as a growth factor, synthesizing inositol phosphate in intercellular locations requiring, but lacking ability to produce or transport adequate quantities of the cell-cell communicator. This idea was experimentally challenged, utilizing a Saccharomyces cerevisiae inositol auxotroph with no MIP enzyme, permeable membranes with a 0.4 mu m pore size, and cellular supernatants as external sources of inositol isolated from S. cerevisiae cells containing either wild-type enzyme (Wt-MIP), no MIP enzyme, auxotroph (Aux), or a green fluorescent protein (GFP) tagged reporter enzyme (MIP- GFP) in co- culturing experiments.ResultsResulting cell densities and microscopic studies with corroborating biochemical and molecular analyses, documented sustained growth of Aux cells in cellular supernatant, concomitant with the uptakeof MIP, detected as MIP-GFP reporter enzyme. These findings revealed previously unknown functions, suggesting that the enzyme can: (1) move into and out of intercellular space, (2) traverse cell walls, and (3) act as a growth factor to promote cellular proliferation of an inositol requiring cell.ConclusionsCo-culturing experiments, designed to test a probable function for MIP secreted in extracellular vesicles, uncovered previously unknown functions for the enzyme and advanced current knowledge concerning spatial control of inositol phosphate biosynthesis. Most importantly, resulting data identified an extracellular vesicle (a non-viral vector) that is capable of synthesizing and transporting inositol phosphate, a biological activity that can be used to enhance specificity of current inositol phosphate therapeutics.en_US
dc.format.mediumelectronic
dc.format.mimetypeapplication/pdf
dc.identifier.citationSteele, E., Alebous, H. D., Vickers, M., Harris, M. E., & Johnson, M. D. (2021). Co-culturing experiments reveal the uptake of myo-inositol phosphate synthase (EC 5.5.1.4) in an inositol auxotroph of Saccharomyces cerevisiae. In Microbial Cell Factories (Vol. 20, Issue 1). Springer Science and Business Media LLC. https://doi.org/10.1186/s12934-021-01610-6
dc.identifier.doi10.1186/s12934-021-01610-6
dc.identifier.urihttps://ir.ua.edu/handle/123456789/11016
dc.languageEnglish
dc.language.isoen_US
dc.publisherBMC
dc.rights.licenseAttribution 4.0 International (CC BY 4.0)
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectInositol auxotroph
dc.subjectSaccharomyces cerevisiae
dc.subjectMyo-inositol Phosphate Synthase (MIP)
dc.subjectProtein secretion
dc.subjectCellular supernatant
dc.subjectExtracellular vesicle
dc.subjectCo-culturing
dc.subjectMYO-INOSITOL-1-PHOSPHATE SYNTHASE
dc.subjectEXTRACELLULAR VESICLES
dc.subjectPROTEIN
dc.subjectLOCALIZATION
dc.subjectBIOSYNTHESIS
dc.subjectYEAST
dc.subjectGENE
dc.subjectEXPRESSION
dc.subjectEXOSOMES
dc.subjectBIOLOGY
dc.subjectBiotechnology & Applied Microbiology
dc.titleCo-culturing experiments reveal the uptake of myo-inositol phosphate synthase (EC 5.5.1.4) in an inositol auxotroph of Saccharomyces cerevisiaeen_US
dc.typeArticle
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