TY - JOUR
T1 - Mass spectrometry-based stable-isotope tracing uncovers metabolic alterations in pyruvate kinase-deficient Aedes aegypti mosquitoes
AU - Petchampai, Natthida
AU - Isoe, Jun
AU - Horvath, Thomas D.
AU - Dagan, Shai
AU - Tan, Lin
AU - Lorenzi, Philip L.
AU - Hawke, David H.
AU - Scaraffia, Patricia Y.
N1 - Funding Information:
This work was financially supported by the Corine Adams Baines Professorship Award, COR Research Bridge Funds Award, U.S. National Institutes of Health , National Institute of Allergy and Infectious Diseases Grant R01AI146199 (to PYS), NIH 1S10OD012304-01 , NIH U01CA235510 , Cancer Prevention and Research Institute of Texas (CPRIT) Grant RP130397 , and The University of Texas MD Anderson's NCI Cancer Center Support Grant P30CA016672 .
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2020/6
Y1 - 2020/6
N2 - A recent in vitro characterization of a recombinant pyruvate kinase (PK) from Aedes aegypti mosquitoes demonstrated that the enzyme is uniquely regulated by multiple allosteric effectors. Here, we further explored PK gene and protein expression, and enzymatic activity in key metabolic tissues of mosquitoes maintained under different nutritional conditions. We also studied the metabolic effects of PK depletion using several techniques including RNA interference and mass spectrometry-based stable-isotope tracing. Transcriptional analysis showed a dynamic post-feeding PK mRNA expression pattern within and across mosquito tissues, whereas corresponding protein levels remained stable throughout the time course analyzed. Nevertheless, PK activity significantly differed in the fat body of sucrose-, blood-fed, and starved mosquitoes. Genetic silencing of PK did not alter survival in blood-fed females maintained on sucrose. However, an enhanced survivorship was observed in PK-deficient females maintained under different nutritional regimens. Our results indicate that mosquitoes overcame PK deficiency by up-regulating the expression of genes encoding NADP-malic enzyme-1, phosphoenolpyruvate carboxykinase-1, phosphoglycerate dehydrogenase and glutamate dehydrogenase, and by decreasing glucose oxidation and metabolic pathways associated with ammonia detoxification. Taken together, our data demonstrate that PK confers to A. aegypti a metabolic plasticity to tightly regulate both carbon and nitrogen metabolism.
AB - A recent in vitro characterization of a recombinant pyruvate kinase (PK) from Aedes aegypti mosquitoes demonstrated that the enzyme is uniquely regulated by multiple allosteric effectors. Here, we further explored PK gene and protein expression, and enzymatic activity in key metabolic tissues of mosquitoes maintained under different nutritional conditions. We also studied the metabolic effects of PK depletion using several techniques including RNA interference and mass spectrometry-based stable-isotope tracing. Transcriptional analysis showed a dynamic post-feeding PK mRNA expression pattern within and across mosquito tissues, whereas corresponding protein levels remained stable throughout the time course analyzed. Nevertheless, PK activity significantly differed in the fat body of sucrose-, blood-fed, and starved mosquitoes. Genetic silencing of PK did not alter survival in blood-fed females maintained on sucrose. However, an enhanced survivorship was observed in PK-deficient females maintained under different nutritional regimens. Our results indicate that mosquitoes overcame PK deficiency by up-regulating the expression of genes encoding NADP-malic enzyme-1, phosphoenolpyruvate carboxykinase-1, phosphoglycerate dehydrogenase and glutamate dehydrogenase, and by decreasing glucose oxidation and metabolic pathways associated with ammonia detoxification. Taken together, our data demonstrate that PK confers to A. aegypti a metabolic plasticity to tightly regulate both carbon and nitrogen metabolism.
KW - Ammonia metabolism
KW - Gene regulation
KW - Mass spectrometry
KW - Metabolomics
KW - Stable-isotope labeled compounds
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U2 - 10.1016/j.ibmb.2020.103366
DO - 10.1016/j.ibmb.2020.103366
M3 - Article
C2 - 32276114
AN - SCOPUS:85083044024
SN - 0965-1748
VL - 121
JO - Insect Biochemistry and Molecular Biology
JF - Insect Biochemistry and Molecular Biology
M1 - 103366
ER -