Knockdown of phosphoethanolamine transmethylation enzymes decreases viability of Haemonchus contortus

William H. Witola, Sheritta Cooks-Fagbodun, Adriana Reyes Ordonez, Kwame Matthews, Daniel A. Abugri, Mark McHugh

Research output: Contribution to journalArticle

Abstract

The phosphobase methylation pathway, in which phosphoethanolamine N-methyltransferases (PMTs) successively catalyze the methylation of phosphoethanolamine to phosphocholine, is essential in the free-living nematode Caenorhabditis elegans. Two PMT-encoding genes (HcPMT1 and HcPMT2) cloned from Haemonchus contortus have been shown, by in vitro assays, to possess enzymatic characteristics similar to those of C. elegans PMTs, but their physiological significance in H. contortus is yet to be elucidated. Therefore, in this study, we endeavored to determine the importance of HcPMT1 and HcPMT2 in the survival of H. contortus by adapting the use of phosphorodiamidate morpholino oligomers (PPMO) antisense approach to block the translation of HcPMT1 and HcPMT2 in the worms. We found that PPMOs targeting HcPMT1 and HcPMT2 down-regulated the expression of HcPMT1 and HcPMT2 proteins in adult H. contortus. Analysis of the effect of HcPMT1 and HcPMT2 knockdown showed that it significantly decreased worm motility and viability, thus validating HcPMT1 and HcPMT2 as essential enzymes for survival of H. contortus. Studies of gene function in H. contortus have been constrained by limited forward and reverse genetic technologies for use in H. contortus. Thus, our success in adaptation of use of PPMO antisense approach in H. contortus provides an important reverse genetic technological advance for studying this parasitic nematode of veterinary significance.

Original languageEnglish (US)
Pages (from-to)1-6
Number of pages6
JournalVeterinary Parasitology
Volume223
DOIs
StatePublished - Jun 15 2016

Fingerprint

Haemonchus
Methyltransferases
Enzymes
Haemonchus contortus
Reverse Genetics
Morpholinos
Caenorhabditis elegans
Methylation
Genes
Phosphorylcholine
Technology
Proteins
In Vitro Techniques
methyltransferases
methylation
viability
enzymes
genes
animal parasitic nematodes
free-living nematodes

Keywords

  • Gene knockdown
  • Haemonchus contortus
  • Phosphoethanolamine N-methyltransferase
  • Phosphorodiamidate morpholino oligomers

ASJC Scopus subject areas

  • Parasitology
  • veterinary(all)

Cite this

Knockdown of phosphoethanolamine transmethylation enzymes decreases viability of Haemonchus contortus. / Witola, William H.; Cooks-Fagbodun, Sheritta; Ordonez, Adriana Reyes; Matthews, Kwame; Abugri, Daniel A.; McHugh, Mark.

In: Veterinary Parasitology, Vol. 223, 15.06.2016, p. 1-6.

Research output: Contribution to journalArticle

Witola WH, Cooks-Fagbodun S, Ordonez AR, Matthews K, Abugri DA, McHugh M. Knockdown of phosphoethanolamine transmethylation enzymes decreases viability of Haemonchus contortus. Veterinary Parasitology. 2016 Jun 15;223:1-6. Available from, DOI: 10.1016/j.vetpar.2016.04.008

Witola, William H.; Cooks-Fagbodun, Sheritta; Ordonez, Adriana Reyes; Matthews, Kwame; Abugri, Daniel A.; McHugh, Mark / Knockdown of phosphoethanolamine transmethylation enzymes decreases viability of Haemonchus contortus.

In: Veterinary Parasitology, Vol. 223, 15.06.2016, p. 1-6.

Research output: Contribution to journalArticle

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abstract = "The phosphobase methylation pathway, in which phosphoethanolamine N-methyltransferases (PMTs) successively catalyze the methylation of phosphoethanolamine to phosphocholine, is essential in the free-living nematode Caenorhabditis elegans. Two PMT-encoding genes (HcPMT1 and HcPMT2) cloned from Haemonchus contortus have been shown, by in vitro assays, to possess enzymatic characteristics similar to those of C. elegans PMTs, but their physiological significance in H. contortus is yet to be elucidated. Therefore, in this study, we endeavored to determine the importance of HcPMT1 and HcPMT2 in the survival of H. contortus by adapting the use of phosphorodiamidate morpholino oligomers (PPMO) antisense approach to block the translation of HcPMT1 and HcPMT2 in the worms. We found that PPMOs targeting HcPMT1 and HcPMT2 down-regulated the expression of HcPMT1 and HcPMT2 proteins in adult H. contortus. Analysis of the effect of HcPMT1 and HcPMT2 knockdown showed that it significantly decreased worm motility and viability, thus validating HcPMT1 and HcPMT2 as essential enzymes for survival of H. contortus. Studies of gene function in H. contortus have been constrained by limited forward and reverse genetic technologies for use in H. contortus. Thus, our success in adaptation of use of PPMO antisense approach in H. contortus provides an important reverse genetic technological advance for studying this parasitic nematode of veterinary significance.",
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