title: Characterization of enzymes from Legionella pneumophila involved in reversible adenylylation of Rab1 protein.
abstract: After the pathogenic bacterium Legionella pneumophila is phagocytosed, it injects more than 250 different proteins into the cytoplasm of host cells to evade lysosomal digestion and to replicate inside the host cell. Among these secreted proteins is the protein DrrA/SidM, which has been shown to modify Rab1b, a main regulator of vesicular trafficking in eukaryotic cells, by transfer of adenosine monophosphate (AMP) to Tyr(77). In addition, Legionella provides the protein SidD that hydrolytically reverses the covalent modification, suggesting a tight spatial and temporal control of Rab1 function by Legionella during infection. Small angle x-ray scattering experiments of DrrA allowed us to validate a tentative complex model built by combining available crystallographic data. We have established the effects of adenylylation on Rab1 interactions and properties in a quantitative way. In addition, we have characterized the kinetics of DrrA-catalyzed adenylylation as well as SidD-catalyzed deadenylylation toward Rab1 and have determined the nucleotide specificities of both enzymes. This study enhances our knowledge of proteins subverting Rab1 function at the Legionella-containing vacuole.
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund.
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Müller
authors.affiliation: European Molecular Biology Laboratory, Hamburg Outstation, EMBL c/o DESY, Notkestrasse 85, Geb 25 A, 22603 Hamburg.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Shkumatov
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund.
authors.forename: Lena K
authors.initials: LK
authors.lastname: Oesterlin
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schoebel
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund.
authors.forename: Philip R
authors.initials: PR
authors.lastname: Goody
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund.
authors.forename: Roger S
authors.initials: RS
authors.lastname: Goody
authors.affiliation: Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund; Center for Integrated Protein Science Munich (CIPSM), Chemistry Department, Technische Universität München, Lichtenbergstrasse 4, 85747 Garching, Germany. Electronic address: aymelt.itzen@tum.de.
authors.forename: Aymelt
authors.initials: A
authors.lastname: Itzen
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016952:Legionella pneumophila / Q000201:enzymology* / Q000235:genetics; D007877:Legionnaires' Disease / Q000201:enzymology* / Q000235:genetics; D011499:Protein Processing, Post-Translational; D020761:rab1 GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D020662:Guanine Nucleotide Exchange Factors; C539733:SidM protein, Legionella pneumophila; C529587:Rab1B protein, human; D020761:rab1 GTP-Binding Proteins