Mitochondrial genes for heme-dependent respiratory chain complexes are up-regulated after depletion of Wolbachia from filarial nematodes.
The filarial nematodes Brugia malayi, Wuchereria bancrofti and Onchocerca volvulus cause elephantiasis or dermatitis and blindness resulting in severe morbidity. Annually, 1.3 billion people are at risk of infection. Targeting the essential Wolbachia endobacteria of filarial nematodes with doxycycline has proven to be an effective therapy resulting in a block in embryogenesis, worm development and macrofilaricidal effects. However, doxycycline is contraindicated for a large portion of the at risk population. To identify new targets for anti-wolbachial therapy, understanding the molecular basis of the Wolbachia-filaria symbiosis is required. Using the B. malayi microarray we identified differentially expressed genes in the rodent filaria Litomosoides sigmodontis after depletion of Wolbachia which might have a role in symbiosis. The microarray data were filtered for regulated genes with a false discovery rate <5% and a > or = 2-fold-change. Most of the genes were differentially expressed at day 36 of tetracycline treatment, when 99.8% of Wolbachia were depleted. Several classes of genes were affected, including genes for translation, transcription, folding/sorting of proteins, motility, structure and metabolic and signalling pathways. Quantitative PCR validated 60% of the genes found to be regulated in the microarray. A nuclear encoded heme-binding protein of the globin family was up-regulated upon loss of Wolbachia. Interestingly, mitochondrial encoded subunits of respiratory chain complexes containing heme and riboflavin were also up-regulated. No change in the expression of these genes was seen in tetracycline treated Wolbachia-free Acanthocheilonema viteae. As Wolbachia synthesise heme and filaria do not, we hypothesise that without the endosymbionts no functional heme-containing enzymes can be formed, leading to loss of energy metabolism which then results in up-regulation of the mitochondrial encoded subunits in an attempt to correct the deviation from homeostasis. Our results support targeting the Wolbachia heme synthesis pathway for the discovery of new anti-filarial drugs.
Strübing
Uta
U
Institute for Medical Microbiology, Immunology and Parasitology, University Clinic Bonn, Sigmund-Freud-Str. 25, D-53105 Bonn, Germany.
Lucius
Richard
R
Hoerauf
Achim
A
Pfarr
Kenneth M
KM
D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D004318:Doxycycline / Q000494:pharmacology; D004579:Electron Transport / Q000235:genetics / Q000502:physiology*; D005370:Filarioidea / Q000235:genetics / Q000378:metabolism*; D050259:Genes, Mitochondrial / Q000235:genetics / Q000502:physiology*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D015854:Up-Regulation; D020577:Wolbachia / Q000187:drug effects / Q000502:physiology*
D000900:Anti-Bacterial Agents; D018076:DNA, Complementary; D004318:Doxycycline