P3 – Characterisation of Fic-enzyme targets from Legionella
Principal Investigators
Prof. Dr. rer. nat. Aymelt Itzen
University Medical Center Hamburg-Eppendorf
Institute of Biochemistry and Signal Transduction
Prof. Dr. Carmen Buchrieser
Institut Pasteur, Paris
Biology of Intracellular Bacteria, Departement Genomes and Genetics
Project Summary
The gram-negative bacterium Legionella pneumophila is the causative agent of Legionnaires’ disease. The phagocytosed pathogen survives in the Legionella containing vacuole (LCV). L. pneumophila secretes a diverse arsenal of about 330 proteins via a Type IV secretion system (T4SS) directly into the macrophage cytosol. These T4SS-effectors manipulate various host processes to downregulate defense mechanisms and promote bacterial survival in the LCV.
A significant fraction of these effectors modifies target proteins by utilizing post-translational modifications (PTMs). For example, the secreted protein SidM/DrrA causes AMPylation in the host’s small GTPase Rab1b. In this project, we will further investigate AMPylations occurring during Legionella infection. We will identify AMPylated host proteins and the respective Legionella enzymes. Using structural biology, infection biology, cell biology and biochemistry, we will uncover the molecular and cellular consequences of AMPylations. The project combines the expertise of Carmen Buchrieser from the Institute Pasteur in understanding Legionella infections with the expertise of Aymelt Itzen in analyzing the impact of PTMs on proteins using structural biology and biochemistry.
References
Du J, Wrisberg M-Kv, Gulen B, Stahl M, Pett C, Hedberg C, Lang K, Schneider S, Itzen A (2021) Rab1-AMPylation by Legionella DrrA is allosterically activated by Rab1. Nat Commun 12:460. Abstract
Hopfner D, Fauser J, Kaspers MS, Pett C, Hedberg C, Itzen A (2020) Monoclonal Anti-AMP Antibodies Are Sensitive and Valuable Tools for Detecting Patterns of AMPylation. iScience 23:101800. Abstract
Gulen B, Rosselin M, Fauser J, Albers MF, Pett C, Krisp C, Pogenberg V, Schluter H, Hedberg C, Itzen A (2020) Identification of targets of AMPylating Fic enzymes by co-substrate-mediated covalent capture. Nat Chem 12:732-9. Abstract
Gomez-Valero L, Rusniok C, Carson D, Mondino S, Perez-Cobas AE, Rolando M, Pasricha S, Reuter S, Demirtas J, Crumbach J, Descorps-Declere S, Hartland EL, Jarraud S, Dougan G, Schroeder GN, Frankel G, Buchrieser C (2019) More than 18,000 effectors in the Legionella genus genome provide multiple, independent combinations for replication in human cells. PNAS 116:2265-73. Abstract
Rolando M, Sanulli S, Rusniok C, Gomez-Valero L, Bertholet C, Sahr T, Margueron R, Buchrieser C (2013) Legionella pneumophila Effector RomA Uniquely Modifies Host Chromatin to Repress Gene Expression and Promote Intracellular Bacterial Replication. Cell Host Microbe 13:395-405. Abstract