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: Research - peer-reviewArticle

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.

LanguageEnglish (US)
Pages1-6
Number of pages6
JournalVeterinary Parasitology
Volume223
DOIs
StatePublished - Jun 15 2016

Fingerprint

Haemonchus
Enzymes
phosphoethanolamine methyltransferase
phosphorylethanolamine
Haemonchus contortus
viability
enzymes
Reverse Genetics
Morpholinos
Caenorhabditis elegans
Methylation
Genes
methyltransferases
Phosphorylcholine
Technology
Proteins
In Vitro Techniques
methylation
genes
animal parasitic 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: Research - peer-reviewArticle

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. 2016 ; Vol. 223. pp. 1-6
@article{2070d1a904484accbeb117dbfae5c337,
title = "Knockdown of phosphoethanolamine transmethylation enzymes decreases viability of Haemonchus contortus",
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.",
keywords = "Gene knockdown, Haemonchus contortus, Phosphoethanolamine N-methyltransferase, Phosphorodiamidate morpholino oligomers",
author = "Witola, {William H.} and Sheritta Cooks-Fagbodun and Ordonez, {Adriana Reyes} and Kwame Matthews and Abugri, {Daniel A.} and Mark McHugh",
year = "2016",
month = "6",
doi = "10.1016/j.vetpar.2016.04.008",
volume = "223",
pages = "1--6",
journal = "Veterinary Parasitology",
issn = "0304-4017",

}

TY - JOUR

T1 - Knockdown of phosphoethanolamine transmethylation enzymes decreases viability of Haemonchus contortus

AU - Witola,William H.

AU - Cooks-Fagbodun,Sheritta

AU - Ordonez,Adriana Reyes

AU - Matthews,Kwame

AU - Abugri,Daniel A.

AU - McHugh,Mark

PY - 2016/6/15

Y1 - 2016/6/15

N2 - 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.

AB - 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.

KW - Gene knockdown

KW - Haemonchus contortus

KW - Phosphoethanolamine N-methyltransferase

KW - Phosphorodiamidate morpholino oligomers

UR - http://www.scopus.com/inward/record.url?scp=84962808824&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84962808824&partnerID=8YFLogxK

U2 - 10.1016/j.vetpar.2016.04.008

DO - 10.1016/j.vetpar.2016.04.008

M3 - Article

VL - 223

SP - 1

EP - 6

JO - Veterinary Parasitology

T2 - Veterinary Parasitology

JF - Veterinary Parasitology

SN - 0304-4017

ER -