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Long-chain acyl-CoA synthetase 2 knockdown leads to decreased fatty acid oxidation in fat body and reduced reproductive capacity in the insect Rhodnius prolixus.
- Source :
-
Biochimica et biophysica acta [Biochim Biophys Acta] 2016 Jul; Vol. 1861 (7), pp. 650-62. Date of Electronic Publication: 2016 Apr 16. - Publication Year :
- 2016
-
Abstract
- Long-chain acyl-CoA esters are important intermediates in lipid metabolism and are synthesized from fatty acids by long-chain acyl-CoA synthetases (ACSL). The hematophagous insect Rhodnius prolixus, a vector of Chagas' disease, produces glycerolipids in the midgut after a blood meal, which are stored as triacylglycerol in the fat body and eggs. We identified twenty acyl-CoA synthetase genes in R. prolixus, two encoding ACSL isoforms (RhoprAcsl1 and RhoprAcsl2). RhoprAcsl1 transcripts increased in posterior midgut on the second day after feeding, and RhoprAcsl2 was highly transcribed on the tenth day. Both enzymes were expressed in Escherichia coli. Recombinant RhoprACSL1 and RhoprACSL2 had broad pH optima (7.5-9.5 and 6.5-9.5, respectively), were inhibited by triacsin C, and were rosiglitazone-insensitive. Both showed similar apparent Km for palmitic and oleic acid (2-6 μM), but different Km for arachidonic acid (0.5 and 6 μM for RhoprACSL1-Flag and RhoprACSL2-Flag, respectively). The knockdown of RhoprAcsl1 did not result in noticeable phenotypes. However, RhoprACSL2 deficient insects exhibited a 2.5-fold increase in triacylglycerol content in the fat body, and 90% decrease in fatty acid β-oxidation. RhoprAcsl2 knockdown also resulted in 20% increase in lifespan, delayed digestion, 30% reduced oviposition, and 50% reduction in egg hatching. Laid eggs and hatched nymphs showed remarkable alterations in morphology. In summary, R. prolixus ACSL isoforms have distinct roles on lipid metabolism. Although RhoprACSL1 functions remain unclear, we propose that RhoprACSL2 is the main contributor for the formation of the intracellular acyl-CoA pool channeled for β-oxidation in the fat body, and is also required for normal reproduction.<br /> (Copyright © 2016 Elsevier B.V. All rights reserved.)
- Subjects :
- Amino Acid Sequence
Animals
Coenzyme A Ligases metabolism
Escherichia coli genetics
Escherichia coli metabolism
Female
Gene Expression Regulation
Gene Knockdown Techniques
Insect Proteins
Isoenzymes genetics
Isoenzymes metabolism
Molecular Sequence Data
Oxidation-Reduction
Phylogeny
Recombinant Proteins genetics
Recombinant Proteins metabolism
Reproduction genetics
Rhodnius classification
Sequence Alignment
Transcription, Genetic
Triazenes
Zygote metabolism
Coenzyme A Ligases genetics
Fat Body metabolism
Fatty Acids metabolism
Oogenesis genetics
Rhodnius genetics
Triglycerides biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 0006-3002
- Volume :
- 1861
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Biochimica et biophysica acta
- Publication Type :
- Academic Journal
- Accession number :
- 27091636
- Full Text :
- https://doi.org/10.1016/j.bbalip.2016.04.007