Structural and functional study of Legionella pneumophila effector RavA.

Publisher: Cold Spring Harbor Laboratory Press Country of Publication: United States NLM ID: 9211750 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1469-896X (Electronic) Linking ISSN: 09618368 NLM ISO Abbreviation: Protein Sci Subsets: MEDLINE

Publication: 2001- : Woodbury, NY : Cold Spring Harbor Laboratory Press
Original Publication: New York, N.Y. : Cambridge University Press, c1992-

Adenosine Triphosphatases/*chemistry ; Adenosine Triphosphatases/*metabolism ; Bacterial Proteins/*chemistry ; Bacterial Proteins/*metabolism ; Legionella pneumophila/*enzymology; Adenosine Triphosphatases/genetics ; Bacterial Proteins/genetics ; HEK293 Cells ; Humans ; Legionella pneumophila/genetics ; Protein Conformation, alpha-Helical ; Protein Domains ; rab GTP-Binding Proteins/chemistry ; rab GTP-Binding Proteins/genetics ; rab GTP-Binding Proteins/metabolism

Legionella pneumophila is an intracellular pathogen that causes Legionnaire's disease in humans. This bacterium can be found in freshwater environments as a free-living organism, but it is also an intracellular parasite of protozoa. Human infection occurs when inhaled aerosolized pathogen comes into contact with the alveolar mucosa and replicates in alveolar macrophages. Legionella enters the host cell by phagocytosis and redirects the Legionella-containing phagosomes from the phagocytic maturation pathway. These nascent phagosomes fuse with ER-derived secretory vesicles and membranes forming the Legionella-containing vacuole. Legionella subverts many host cellular processes by secreting over 300 effector proteins into the host cell via the Dot/Icm type IV secretion system. The cellular function for many Dot/Icm effectors is still unknown. Here, we present a structural and functional study of L. pneumophila effector RavA (Lpg0008). Structural analysis revealed that the RavA consists of four ~85 residue long α-helical domains with similar folds, which show only a low level of structural similarity to other protein domains. The ~90 residues long C-terminal segment is predicted to be natively unfolded. We show that during L. pneumophila infection of human cells, RavA localizes to the Golgi apparatus and to the plasma membrane. The same localization is observed when RavA is expressed in human cells. The localization signal resides within the C-terminal sequence C ⁴⁰⁹ WTSFCGLF ⁴¹⁷ . Yeast-two-hybrid screen using RavA as bait identified RAB11A as a potential binding partner. RavA is present in L. pneumophila strains but only distant homologs are found in other Legionella species, where the number of repeats varies.
(© 2021 The Protein Society.)

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MOP 48370 Canada CIHR

Keywords: Golgi localization; Golgi localization sequence; Legionella pneumophila; crystal structure; domain structure; effector RavA

0 (Bacterial Proteins)
EC 3.6.1.- (Adenosine Triphosphatases)
EC 3.6.1.- (rab11 protein)
EC 3.6.5.2 (rab GTP-Binding Proteins)

Date Created: 20210304 Date Completed: 20211223 Latest Revision: 20220503

20221213

PMC8040872

10.1002/pro.4057

33660322