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Using Euf1 transcription factor as a titrator of erythritol-inducible promoters in Yarrowia lipolytica; insight into the structure, splicing, and regulation mechanism

cris.lastimport.scopus2025-10-23T06:56:01Z
cris.virtual.author-orcid0000-0001-8372-8459
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cris.virtualsource.author-orcid6f5a4155-2edb-48cf-b362-f3180e151169
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dc.abstract.enControllable regulatory elements, like inducible, titratable promoters, are highly desired in synthetic biology toolboxes. A set of previously developed erythritol-inducible promoters along with an engineered Yarrowia lipolytica host strain were shown to be a very potent expression platform. In this study, we push the previously encountered limits of the synthetic promoters’ titratability (by the number of upstream motifs) by using a compatible transcription factor, Euf1, as the promoter titrator. Overexpression of spliced EUF1 turned out to be very efficient in promoting expression from the compatible promoter, however, the erythritol-inducible character of the promoter was then lost. Analysis of the EUF1’s splicing pattern suggests that the intron removal is promoted in the presence of erythritol, but is not dependent on it. The 3D structures of spliced versus unspliced Euf1 were modeled, and ligand-binding strength was calculated and compared. Furthermore, the EUF1-dependent expression profile under different chemical stimulants was investigated. Depletion of carbon source was identified as the significant factor upregulating the expression from the Euf1-dependent promoter (2–10-fold). Considering these findings and transcriptomics data, a new mechanism of the Euf1-regulated promoter action is proposed, involving a ‘catabolite repression’ transcription factor—Adr1, both acting on the same ERY-inducible promoter.
dc.affiliationWydział Nauk o Żywności i Żywieniu
dc.affiliation.instituteKatedra Biotechnologii i Mikrobiologii Żywności
dc.contributor.authorCelińska, Ewelina
dc.contributor.authorKorpys-Woźniak, Paulina
dc.contributor.authorGorczyca, Maria
dc.contributor.authorNicaud, Jean-Marc
dc.date.access2025-07-22
dc.date.accessioned2025-07-22T08:17:46Z
dc.date.available2025-07-22T08:17:46Z
dc.date.copyright2024-08-21
dc.date.issued2024
dc.description.abstract<jats:title>Abstract</jats:title> <jats:p>Controllable regulatory elements, like inducible, titratable promoters, are highly desired in synthetic biology toolboxes. A set of previously developed erythritol-inducible promoters along with an engineered Yarrowia lipolytica host strain were shown to be a very potent expression platform. In this study, we push the previously encountered limits of the synthetic promoters’ titratability (by the number of upstream motifs) by using a compatible transcription factor, Euf1, as the promoter titrator. Overexpression of spliced EUF1 turned out to be very efficient in promoting expression from the compatible promoter, however, the erythritol-inducible character of the promoter was then lost. Analysis of the EUF1’s splicing pattern suggests that the intron removal is promoted in the presence of erythritol, but is not dependent on it. The 3D structures of spliced versus unspliced Euf1 were modeled, and ligand-binding strength was calculated and compared. Furthermore, the EUF1-dependent expression profile under different chemical stimulants was investigated. Depletion of carbon source was identified as the significant factor upregulating the expression from the Euf1-dependent promoter (2–10-fold). Considering these findings and transcriptomics data, a new mechanism of the Euf1-regulated promoter action is proposed, involving a ‘catabolite repression’ transcription factor—Adr1, both acting on the same ERY-inducible promoter.</jats:p>
dc.description.accesstimeat_publication
dc.description.bibliographyil., bibliogr.
dc.description.financepublication_nocost
dc.description.financecost0,00
dc.description.if2,7
dc.description.points100
dc.description.versionfinal_published
dc.description.volume24
dc.identifier.doi10.1093/femsyr/foae027
dc.identifier.eissn1567-1364
dc.identifier.issn1567-1356
dc.identifier.urihttps://sciencerep.up.poznan.pl/handle/item/3916
dc.identifier.weblinkhttps://academic.oup.com/femsyr/article/doi/10.1093/femsyr/foae027/7738376?login=true
dc.languageen
dc.relation.ispartofFEMS Yeast Research
dc.relation.pagesfoae027
dc.relation.project2021/41/B/NZ9/00086
dc.rightsCC-BY-NC
dc.sciencecloudsend
dc.share.typeOPEN_JOURNAL
dc.subject.enrecombinant protein
dc.subject.enheterologous expression
dc.subject.ensynthetic promoter
dc.subject.eninducible expression
dc.subject.enyeast
dc.subject.ensplicing regulation
dc.titleUsing Euf1 transcription factor as a titrator of erythritol-inducible promoters in Yarrowia lipolytica; insight into the structure, splicing, and regulation mechanism
dc.typeJournalArticle
dspace.entity.typePublication
oaire.citation.volume24
project.funder.name2021/41/B/NZ9/00086