Listeria monocytogenes genes supporting growth under standard laboratory cultivation conditions and during macrophage infection [RESEARCH]

Martin A. Fischer1,4, Tim Engelgeh1, Patricia Rothe1, Stephan Fuchs2, Andrea Thürmer3 and Sven Halbedel1 1FG11 Division of Enteropathogenic Bacteria and Legionella, Robert Koch Institute, 38855 Wernigerode, Germany; 2MF1 Bioinformatic Support, Robert Koch Institute, 13353 Berlin, Germany; 3MF2 Genome Sequencing, Robert Koch Institute, 13353 Berlin, Germany

4 Present address: FG13 Nosocomial Pathogens and Antibiotic Resistances, Robert Koch Institute, 38855 Wernigerode, Germany

Corresponding author: halbedelsrki.de Abstract

The Gram-positive bacterium Listeria monocytogenes occurs widespread in the environment and infects humans when ingested along with contaminated food. Such infections are particularly dangerous for risk group patients, for whom they represent a life-threatening disease. To invent novel strategies to control contamination and disease, it is important to identify those cellular processes that maintain pathogen growth inside and outside the host. Here, we have applied transposon insertion sequencing (Tn-Seq) to L. monocytogenes for the identification of such processes on a genome-wide scale. Our approach identified 394 open reading frames that are required for growth under standard laboratory conditions and 42 further genes, which become necessary during intracellular growth in macrophages. Most of these genes encode components of the translation machinery and act in chromosome-related processes, cell division, and biosynthesis of the cellular envelope. Several cofactor biosynthesis pathways and 29 genes with unknown functions are also required for growth, suggesting novel options for the development of antilisterial drugs. Among the genes specifically required during intracellular growth are known virulence factors, genes compensating intracellular auxotrophies, and several cell division genes. Our experiments also highlight the importance of PASTA kinase signaling for general viability and of glycine metabolism and chromosome segregation for efficient intracellular growth of L. monocytogenes.

Received March 11, 2022. Accepted August 4, 2022.

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