A naturally occurring prfA truncation in a Listeria monocytogenes field strain contributes to reduced replication and cell-to-cell spread

Rupp, Sebastian; Aguilar Bultet, Lisandra; Jagannathan, Vidhya; Guldimann, Claudia; Drögemüller, Cord; Pfarrer, Christiane; Vidondo, Beatriz; Seuberlich, Torsten; Frey, Joachim; Oevermann, Anna (2015). A naturally occurring prfA truncation in a Listeria monocytogenes field strain contributes to reduced replication and cell-to-cell spread. Veterinary microbiology, 179(1-2), pp. 91-101. Elsevier 10.1016/j.vetmic.2015.03.002

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Listeria (L.) monocytogenes is an environmental bacterium that may become an intracellular pathogen upon ingestion to cause gastroenteritis, septicaemia, abortions, and/or fatal infections of the central nervous system. We here describe a L. monocytogenes field strain (JF5171) isolated from a bovine placenta in the context of abortion, which exhibited attenuation in bovine brain-slice cultures. The whole genome of strain JF5171 was sequenced, and the invasion, replication, and intercellular spread of JF5171 were further analyzed by quantification of colony forming units and immunofluorescence studies. Phospholipase and hemolysis activity of JF5171 were also quantified along with transcription levels of actA, hly and prfA. The data obtained were compared to those of the widely used L. monocytogenes reference strain, EGD-e. JF5171 exhibited reduced replication and lower levels of phospholipase and hemolysis activity. Invasion and cell-to-cell spread was strongly decreased compared to EGD-e, and actin polymerization was absent. A frame shift deletion was identified in the JF5171 coding region of the major regulator for virulence, prfA. This resulted in a truncated C-terminus sequence (WEN* vs. WGKLN*). In addition, a point mutation resulted in a lysine to arginine substitution at amino acid position 197. Complementation with prfA from EGD-e and with (EGD-e) prfA-K197N increased the replication and spread efficiency of JF5171. In contrast, complementation with the truncated version of prfA had no effect. Taken together, these results suggest that the truncated C-terminus of prfA considerably contributes to the strongly attenuated phenotype observed in vitro.

Item Type:

Journal Article (Original Article)

Division/Institute:

05 Veterinary Medicine > Research Foci > NeuroCenter
05 Veterinary Medicine > Research Foci > Veterinary Public Health / Herd Health Management
05 Veterinary Medicine > Research Foci > Host-Pathogen Interaction
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Veterinary Public Health Institute
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Experimental Clinical Research
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH) > Institute of Genetics
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP)
05 Veterinary Medicine > Department of Clinical Research and Veterinary Public Health (DCR-VPH)
05 Veterinary Medicine > Department of Infectious Diseases and Pathobiology (DIP) > Institute of Veterinary Bacteriology

Graduate School:

Graduate School for Cellular and Biomedical Sciences (GCB)

UniBE Contributor:

Rupp, Sebastian; Aguilar Bultet, Lisandra; Drögemüller, Cord; Vidondo Curras, Beatriz; Seuberlich, Torsten; Frey, Joachim and Oevermann, Anna

Subjects:

500 Science > 570 Life sciences; biology

ISSN:

0378-1135

Publisher:

Elsevier

Funders:

[4] Swiss National Science Foundation
[8] 3R Research Foundation Switzerland

Language:

English

Submitter:

Sebastian Rupp

Date Deposited:

19 Jun 2015 09:46

Last Modified:

11 Oct 2018 17:01

Publisher DOI:

10.1016/j.vetmic.2015.03.002

PubMed ID:

25813546

BORIS DOI:

10.7892/boris.69640

URI:

https://boris.unibe.ch/id/eprint/69640

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