The KdpD/KdpE two-component system of Photorhabdus asymbiotica promotes bacterial survival within M. sexta hemocytes

Isabella Vlisidou, I Eleftherianos, Stephen Dorus, Guowei Yang, R H Ffrench-Constant, Stuart E Reynolds, Nicholas R Waterfield

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Many bacteria persist within phagocytes, deploying complex sets of tightly regulated virulence factors to manipulate and survive within host cells. So far, no single factor has been identified that is sufficient to allow intracellular persistence of an otherwise non-pathogenic bacterium. Here we report that the two-component KdpD/KdpE sensor kinase/response regulator of the insect and human pathogen Photorhabdus asymbiotica (Pa) is sufficient to allow a harmless laboratory strain of E. coil to resist phagocytic killing and persist within insect hemocytes, ultimately killing the insect. Screening of a cosmid library of Pa in E. coil by injection into the moth Manduca sexta, previously identified three overlapping clones which caused the insect to cease feeding and subsequently die. Transposon mutagenesis revealed a cosmid encoded kdp high affinity potassium pump regulon was responsible for this phenotype. Gentamycin protection assays and confocal microscopy revealed the cosmid clones were persisting inside insect hemocytes far longer than control bacteria. Cloning and expression of Pa kdpD/kdpE alone into E. coil recapitulated the phenotype. Bioassay results and transcriptional analysis of various E. coil kdp mutants harboring the Pa kdp genes confirmed that Pa KdpD/KdpE was able to induce the E. coil kdp pump structural genes in response to exposure to insect hemocytes but not blood plasma alone. The finding that Pa KdpD/KdpE can facilitate resistance of E. colt to phagocytic killing suggests a central role for potassium in this process, supporting previous work implicating potassium sensing in virulence of other bacteria and also in the normal process of protease killing of engulfed bacteria by neutrophils.
Original languageEnglish
Pages (from-to)352-362
Number of pages11
JournalJournal of Invertebrate Pathology
Volume105
Issue number3
DOIs
Publication statusPublished - Nov 2010

Fingerprint

hemocytes
insect
insects
bacterium
bacteria
potassium
virulence
phenotype
clone
pump
clones
regulon
colts
sodium-potassium-exchanging ATPase
gene
structural genes
phagocytes
Manduca sexta
gentamicin
blood plasma

Keywords

  • photorhabdus
  • hemocytes
  • Kdp potassium pump
  • phagocytosis

Cite this

The KdpD/KdpE two-component system of Photorhabdus asymbiotica promotes bacterial survival within M. sexta hemocytes. / Vlisidou, Isabella; Eleftherianos, I; Dorus, Stephen; Yang, Guowei; Ffrench-Constant, R H; Reynolds, Stuart E; Waterfield, Nicholas R.

In: Journal of Invertebrate Pathology, Vol. 105, No. 3, 11.2010, p. 352-362.

Research output: Contribution to journalArticle

Vlisidou, Isabella ; Eleftherianos, I ; Dorus, Stephen ; Yang, Guowei ; Ffrench-Constant, R H ; Reynolds, Stuart E ; Waterfield, Nicholas R. / The KdpD/KdpE two-component system of Photorhabdus asymbiotica promotes bacterial survival within M. sexta hemocytes. In: Journal of Invertebrate Pathology. 2010 ; Vol. 105, No. 3. pp. 352-362.
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