| Measurement of effector protein injection by type III and type IV secretion systems by using a 13-residue phosphorylatable glycogen synthase kinase tag. | |
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MedLine Citation:
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PMID: 16988240 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Numerous bacterial pathogens use type III secretion systems (T3SSs) or T4SSs to inject or translocate virulence proteins into eukaryotic cells. Several different reporter systems have been developed to measure the translocation of these proteins. In this study, a peptide tag-based reporter system was developed and used to monitor the injection of T3S and T4S substrates. The glycogen synthase kinase (GSK) tag is a 13-residue phosphorylatable peptide tag derived from the human GSK-3beta kinase. Translocation of a GSK-tagged protein into a eukaryotic cell results in host cell protein kinase-dependent phosphorylation of the tag, which can be detected with phosphospecific GSK-3beta antibodies. A series of expression plasmids encoding Yop-GSK fusion proteins were constructed to evaluate the ability of the GSK tag to measure the injection of Yops by the Yersinia pestis T3SS. GSK-tagged YopE, YopH, LcrQ, YopK, YopN, and YopJ were efficiently phosphorylated when translocated into HeLa cells. Similarly, the injection of GSK-CagA by the Helicobacter pylori T4SS into different cell types was measured via phosphorylation of the GSK tag. The GSK tag provides a simple method to monitor the translocation of T3S and T4S substrates. |
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Authors:
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Julie Torruellas Garcia; Franco Ferracci; Michael W Jackson; Sabrina S Joseph; Isabelle Pattis; Lisa R W Plano; Wolfgang Fischer; Gregory V Plano |
Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural |
Journal Detail:
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Title: Infection and immunity Volume: 74 ISSN: 0019-9567 ISO Abbreviation: Infect. Immun. Publication Date: 2006 Oct |
Date Detail:
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Created Date: 2006-09-21 Completed Date: 2006-11-28 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 0246127 Medline TA: Infect Immun Country: United States |
Other Details:
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Languages: eng Pagination: 5645-57 Citation Subset: IM |
Affiliation:
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Department of Microbiology and Immunology, University of Miami Miller School of Medicine, P.O. Box 016960 (R-138), Miami, FL 33101, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Bacterial Outer Membrane Proteins / genetics, metabolism* Bacterial Proteins / genetics, metabolism* Genetic Vectors / genetics Glycogen Synthase Kinases / analysis*, genetics, metabolism* Hela Cells Helicobacter pylori / metabolism Humans Molecular Sequence Data Phosphorylation Plasmids / genetics Protein Transport Recombinant Fusion Proteins / genetics, metabolism Virulence Factors / analysis*, genetics, metabolism* Yersinia pestis / metabolism |
| Grant Support | |
ID/Acronym/Agency:
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AI-39575/AI/NIAID NIH HHS |
| Chemical | |
Reg. No./Substance:
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0/Bacterial Outer Membrane Proteins; 0/Bacterial Proteins; 0/Recombinant Fusion Proteins; 0/Virulence Factors; EC 2.7.11.-/Glycogen Synthase Kinases |
| Comments/Corrections | |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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