Umeå University's logo

umu.sePublications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
V. cholerae MakA is a cholesterol-binding pore-forming toxin that induces non-canonical autophagy
Umeå University, Faculty of Science and Technology, Department of Chemistry. Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).ORCID iD: 0000-0002-1700-3668
Umeå University, Faculty of Science and Technology, Department of Chemistry. Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
Umeå University, Faculty of Science and Technology, Department of Chemistry. Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
Umeå University, Faculty of Science and Technology, Department of Chemistry. Umeå University, Faculty of Medicine, Umeå Centre for Microbial Research (UCMR).
Show others and affiliations
2022 (English)In: Journal of Cell Biology, ISSN 0021-9525, E-ISSN 1540-8140, Vol. 221, no 12, article id e202206040Article in journal (Refereed) Published
Abstract [en]

Pore-forming toxins (PFTs) are important virulence factors produced by many pathogenic bacteria. Here, we show that the Vibrio cholerae toxin MakA is a novel cholesterol-binding PFT that induces non-canonical autophagy in a pH-dependent manner. MakA specifically binds to cholesterol on the membrane at pH < 7. Cholesterol-binding leads to oligomerization of MakA on the membrane and pore formation at pH 5.5. Unlike other cholesterol-dependent cytolysins (CDCs) which bind cholesterol through a conserved cholesterol-binding motif (Thr-Leu pair), MakA contains an Ile-Ile pair that is essential for MakA-cholesterol interaction. Following internalization, endosomal acidification triggers MakA pore-assembly followed by ESCRT-mediated membrane repair and V-ATPase-dependent unconventional LC3 lipidation on the damaged endolysosomal membranes. These findings characterize a new cholesterol-binding toxin that forms pores in a pH-dependent manner and reveals the molecular mechanism of host autophagy manipulation.

Place, publisher, year, edition, pages
Rockefeller University Press, 2022. Vol. 221, no 12, article id e202206040
Keywords [en]
cholesterol-binding, MakA, non-canonical autophagy, pore-forming toxin, Vibrio Cholerae
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-200014DOI: 10.1083/jcb.202206040ISI: 000932911400001PubMedID: 36194176Scopus ID: 2-s2.0-85139366240OAI: oai:DiVA.org:umu-200014DiVA, id: diva2:1701387
Funder
Knut and Alice Wallenberg FoundationEU, European Research CouncilSwedish Research Council, 2018-04585Göran Gustafsson Foundation for Research in Natural Sciences and MedicineAvailable from: 2022-10-05 Created: 2022-10-05 Last updated: 2024-03-27Bibliographically approved
In thesis
1. Non-canonical ATG8 conjugation in ESCRT-driven membrane remodeling processes
Open this publication in new window or tab >>Non-canonical ATG8 conjugation in ESCRT-driven membrane remodeling processes
2024 (English)Doctoral thesis, comprehensive summary (Other academic)
Alternative title[sv]
Okonventionell ATG8-konjugering i ESCRT-drivna membranombyggnadsprocesser
Abstract [en]

ATG8 family proteins have the unique ability to conjugate to membrane lipids. Initially identified as a hallmark of autophagy, ATG8 lipidation is emerging as an important regulator of a growing list of non-degradative cellular functions. In this thesis we developed and applied novel chemical genetic approaches to perturb dynamic membrane remodeling processes and induce non-canonical ATG8 conjugation in cells. We investigated novel roles of ATG8 in membrane deformation processes carried out by the Endosomal Sorting Complex Requiredfor Transport (ESCRT) machinery.

In Paper I, using a high-throughput phenotypic screening assay, we developed a collection of pseudo-natural product-based compounds which potently induce ATG8 lipidation in mammalian cells. The most potent compound, Tantalosin, induces ATG8 lipidation which is insensitive to simultaneous inhibition of autophagosome-lysosome fusion, suggesting a non-canonical function ofTantalosin-induced ATG8 conjugation.

In Paper II we investigated the molecular target of Tantalosin. We found that Tantalosin targets the ESCRT-III protein IST1 and inhibits IST1-CHMP1B copolymer formation. This inhibition results in the impairment of transferrin receptor (TfR) recycling resulting in the rapid accumulation of the receptor inearly/sorting endosomes. At the same time, Tantalosin induces non-canonical ATG8 conjugation on stalled sorting endosomes containing TfR. This conjugation is dependent on the ATG16L1-ATG5-ATG12 complex which is recruited to stalled endosomes via ATG16L1-V-ATPase interaction.

In Paper III and Paper IV we studied the induction of non-canonical ATG8 lipidation in response to endolysosomal membrane damage. We used two established membrane damaging agents: V. Cholerae cytotoxin MakA and the lysosomotropic compound, LLOMe. In Paper III we demonstrated that, at lowpH, MakA assembles into small pores in endosomal membranes which arerecognized by the ESCRT membrane repair machinery. Non-canonical ATG8 lipidation in response to MakA-induced pore formation is mediated by V-ATPase activity. In Paper IV we identified a novel player in the lysosomal damage response – TECRP1. TECPR1 is recruited to damaged membranes where it forms an alternative ATG16L1-independent E3 ligase complex with the ATG5-ATG12 conjugate and plays a role in the restoration of lysosomal integrity after damage.

Place, publisher, year, edition, pages
Umeå: Umeå University, 2024. p. 70
Keywords
ATG8 conjugation, Endosomal Sorting Complex Required for Transport, membrane remodeling, chemical genetics
National Category
Cell Biology Biochemistry and Molecular Biology
Identifiers
urn:nbn:se:umu:diva-222756 (URN)978-91-8070-356-7 (ISBN)978-91-8070-357-4 (ISBN)
Public defence
2024-04-26, Stora Hörsalen, KBC byggnad KBE303, 09:00 (English)
Opponent
Supervisors
Available from: 2024-04-05 Created: 2024-03-27 Last updated: 2024-03-27Bibliographically approved

Open Access in DiVA

fulltext(6429 kB)168 downloads
File information
File name FULLTEXT01.pdfFile size 6429 kBChecksum SHA-512
39b1ba6d2bf353a3f1f3a02f4a9c685ac5be5c79db4fa87c14af6a303c8508480b6b93641a5eedd1d46d55845f768a1bf91d4e99532692a47f4ebe2726d2ac60
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMedScopus

Authority records

Jia, XiaotongKnyazeva, AnastasiaZhang, YuCastro-Gonzalez, SergioCarlson, Lars-AndersCorkery, DaleWu, Yao-Wen

Search in DiVA

By author/editor
Jia, XiaotongKnyazeva, AnastasiaZhang, YuCastro-Gonzalez, SergioCarlson, Lars-AndersYoshimori, TamotsuCorkery, DaleWu, Yao-Wen
By organisation
Department of ChemistryUmeå Centre for Microbial Research (UCMR)Department of Medical Biochemistry and BiophysicsMolecular Infection Medicine Sweden (MIMS)Wallenberg Centre for Molecular Medicine at Umeå University (WCMM)
In the same journal
Journal of Cell Biology
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 168 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 695 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf