An acetyltransferase effector conserved across Legionella species targets the eukaryotic eIF3 complex to modulate protein translation

• Published in: mBio 2024-03, Vol.15 (3), p.e0322123-e0322123
• Main Authors: Syriste, Lukas, Patel, Deepak T, Stogios, Peter J, Skarina, Tatiana, Patel, Dhruvin, Savchenko, Alexei
• Format: Article
• Language: English
• Subjects: acetyltransferase; Acetyltransferases - metabolism; Bacterial Proteins - metabolism; effectors; Eukaryotic Initiation Factor-3 - metabolism; Host-Microbial Interactions; Humans; Legionella - genetics; Legionella pneumophila; Legionella pneumophila - genetics; Legionnaires' Disease; Lysine - metabolism; pathogenesis; Prokaryotic Initiation Factor-3 - metabolism; Protein Biosynthesis; Research Article
• ISSN: 2150-7511; 2150-7511
• DOI: 10.1128/mbio.03221-23

The survival of spp. as intracellular pathogens relies on the combined action of protein effectors delivered inside their eukaryotic hosts by the Dot/Icm ( efective in rganelle rafficking/ ntra ellular ultiplication) type IVb secretion system. The specific repertoire of effector arsenals varies dramatically across over 60 known species of this genera with responsible for most cases of Legionnaires' disease in humans encoding over 360 Dot/Icm effectors. However, a small subset of "core" effectors appears to be conserved across all species raising an intriguing question of their role in these bacteria's pathogenic strategy, which for most of these effectors remains unknown. Lpg0103 effector, also known as VipF, represents one of the core effector families that features a tandem of Gcn5-related N-acetyltransferase (GNAT) domains. Here, we present the crystal structure of the Lha0223, the VipF representative from in complex with acetyl-coenzyme A determined to 1.75 Å resolution. Our structural analysis suggested that this effector family shares a common fold with the two GNAT domains forming a deep groove occupied by residues conserved across VipF homologs. Further analysis suggested that only the C-terminal GNAT domain of VipF effectors retains the active site composition compatible with catalysis, whereas the N-terminal GNAT domain binds the ligand in a non-catalytical mode. We confirmed this by enzymatic assays which revealed VipF activity not only against generic small molecule substrates, such as chloramphenicol, but also against poly-L-lysine and histone-derived peptides. We identified the human eukaryotic translation initiation factor 3 (eIF3) complex co-precipitating with Lpg0103 and demonstrated the direct interaction between the several representatives of the VipF family, including Lpg0103 and Lha0223 with the K subunit of eIF3. According to our data, these interactions involve primarily the C-terminal tail of eIF3-K containing two lysine residues that are acetylated by VipF. VipF catalytic activity results in the suppression of eukaryotic protein translation , revealing the potential function of VipF "core" effectors in 's pathogenic strategy.IMPORTANCEBy translocating effectors inside the eukaryotic host cell, bacteria can modulate host cellular processes in their favor. species, which includes the pneumonia-causing encode a widely diverse set of effectors with only a small subset that is conserved across this genus. Here, we demonstrate that one o