uu.seUppsala University Publications
Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • 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
An experimental pipeline for initial characterization of bacterial type III secretion system inhibitor mode of action using enteropathogenic Yersinia
Show others and affiliations
2018 (English)In: Frontiers in Cellular and Infection Microbiology, E-ISSN 2235-2988, Vol. 8, article id 404Article in journal (Refereed) Published
Abstract [en]

Dozens of Gram negative pathogens use one or more type III secretion systems (T3SS) to disarm host defenses or occupy a beneficial niche during infection of a host organism. While the T3SS represents an attractive drug target and dozens of compounds with T3SS inhibitory activity have been identified, few T3SS inhibitors have been validated and mode of action determined. One issue is the lack of standardized orthogonal assays following high throughput screening. Using a training set of commercially available compounds previously shown to possess T3SS inhibitory activity, we demonstrate the utility of an experiment pipeline comprised of six distinct assays to assess the stages of type III secretion impacted: T3SS gene copy number, T3SS gene expression, T3SS basal body and needle assembly, secretion of cargo through the T3SS, and translocation of T3SS effector proteins into host cells. We used enteropathogenic Yersinia as the workhorse T3SS-expressing model organisms for this experimental pipeline, as Yersinia is sensitive to all T3SS inhibitors we tested, including those active against other T3SS-expressing pathogens. We find that this experimental pipeline is capable of rapidly distinguishing between T3SS inhibitors that interrupt the process of type III secretion at different points in T3SS assembly and function. For example, our data suggests that Compound 3, a malic diamide, blocks either activity of the assembled T3SS or alters the structure of the T3SS in a way that blocks T3SS cargo secretion but not antibody recognition of the T3SS needle. In contrast, our data predicts that Compound 4, a haloid-containing sulfonamidobenzamide, disrupts T3SS needle subunit secretion or assembly. Furthermore, we suggest that misregulation of copy number control of the pYV virulence plasmid, which encodes the Yersinia T3SS, should be considered as a possible mode of action for compounds with T3SS inhibitory activity against Yersinia.

Place, publisher, year, edition, pages
2018. Vol. 8, article id 404
National Category
Medicinal Chemistry Microbiology
Research subject
Microbiology
Identifiers
URN: urn:nbn:se:uu:diva-366601DOI: 10.3389/fcimb.2018.00404ISI: 000450941000001PubMedID: 30524970OAI: oai:DiVA.org:uu-366601DiVA, id: diva2:1265053
Available from: 2018-11-22 Created: 2018-11-22 Last updated: 2019-01-22Bibliographically approved

Open Access in DiVA

fulltext(3346 kB)88 downloads
File information
File name FULLTEXT01.pdfFile size 3346 kBChecksum SHA-512
9e9b573c2e59863be2b8e8a65b5ee60d2ab6ba49be72c2aafe111552a5b42084f1430221d8d921d6c347954dad0a58a05a6a3c1af9002591ec94c2105b14984f
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed

Authority records BETA

Wang, Helen

Search in DiVA

By author/editor
Wang, Helen
By organisation
Department of Medical Biochemistry and Microbiology
In the same journal
Frontiers in Cellular and Infection Microbiology
Medicinal ChemistryMicrobiology

Search outside of DiVA

GoogleGoogle Scholar
Total: 88 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: 124 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association
  • 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