Subcellular localization of legionella Dot/Icm effectors

Adam J. Vogrin, Aurelie Mousnier, Gad Frankel, Elizabeth L. Hartland

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Researchpeer-review

Abstract

The translocation of effector proteins by the Dot/Icm type IV secretion system is central to the ability of Legionella pneumophila to persist and replicate within eukaryotic cells. The subcellular localization of translocated Dot/Icm proteins in host cells provides insight into their function. Through co-staining with host cell markers, effector proteins may be localized to specific subcellular compartments and membranes, which frequently reflects their host cell target and mechanism of action. In this chapter, we describe protocols to (1) localize effector proteins within cells by ectopic expression using green fluorescent protein fusions and (2) localize effector proteins within infected cells using epitope-tagged effector proteins and immuno-fluorescence microscopy.

Original languageEnglish
Title of host publicationLegionella: Methods and Protocols
PublisherHumana Press
Pages333-344
Number of pages12
Volume954
ISBN (Print)9781627031608
DOIs
Publication statusPublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume954
ISSN (Print)10643745

Keywords

  • Epitope tagging
  • HEK293T cells
  • Infection
  • Laser scanning confocal microscopy
  • Legionella pneumophila
  • Transfection
  • Type IV secretion

Cite this

Vogrin, A. J., Mousnier, A., Frankel, G., & Hartland, E. L. (2013). Subcellular localization of legionella Dot/Icm effectors. In Legionella: Methods and Protocols (Vol. 954, pp. 333-344). (Methods in Molecular Biology; Vol. 954). Humana Press. https://doi.org/10.1007/978-1-62703-161-5-20