Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2020 Dec 15:9:e58053.
doi: 10.7554/eLife.58053.

Downregulation of the tyrosine degradation pathway extends Drosophila lifespan

Andrey A Parkhitko  1   2 , Divya Ramesh  3   4 , Lin Wang  5   6 , Dmitry Leshchiner  1 , Elizabeth Filine  1 , Richard Binari  1   7 , Abby L Olsen  8 , John M Asara  9 , Valentin Cracan  10   11 , Joshua D Rabinowitz  5   6 , Axel Brockmann  3 , Norbert Perrimon  1   7
Affiliations

Downregulation of the tyrosine degradation pathway extends Drosophila lifespan

Andrey A Parkhitko et al. Elife. .

Abstract

Aging is characterized by extensive metabolic reprogramming. To identify metabolic pathways associated with aging, we analyzed age-dependent changes in the metabolomes of long-lived Drosophila melanogaster. Among the metabolites that changed, levels of tyrosine were increased with age in long-lived flies. We demonstrate that the levels of enzymes in the tyrosine degradation pathway increase with age in wild-type flies. Whole-body and neuronal-specific downregulation of enzymes in the tyrosine degradation pathway significantly extends Drosophila lifespan, causes alterations of metabolites associated with increased lifespan, and upregulates the levels of tyrosine-derived neuromediators. Moreover, feeding wild-type flies with tyrosine increased their lifespan. Mechanistically, we show that suppression of ETC complex I drives the upregulation of enzymes in the tyrosine degradation pathway, an effect that can be rescued by tigecycline, an FDA-approved drug that specifically suppresses mitochondrial translation. In addition, tyrosine supplementation partially rescued lifespan of flies with ETC complex I suppression. Altogether, our study highlights the tyrosine degradation pathway as a regulator of longevity.

Keywords: D. melanogaster; ETC Complex I; TAT; genetics; genomics; mitochondria; neurotransmitters; tigecycline; tyrosine aminotransferase.

PubMed Disclaimer

Conflict of interest statement

AP, DR, LW, DL, EF, RB, AO, JA, VC, JR, AB, NP No competing interests declared

Figures

Figure 1.
Figure 1.. Tyrosine is a new lifespan-dependent metabolite.
(A) Heat map showing the metabolites that significantly changed in 1-week and 4-week-old wild-type (B3) and long-lived (O1 and O3) flies. Each row represents a mean of five biological replicates. (B) Box plots of relative levels of tyrosine in 1-week and 4-week-old wild-type (B3) and long-lived (O1 and O3) flies extracted from the heat map (A). (C) Relative mRNA levels of CG1461 in 1-week-old control (B3) and long-lived (O1, O3) flies. Means ± SD. (D) Relative mRNA levels of CG1461, Faa, Hgo, CG11796, GstZ2, and Hn from 1-week and 5-week-old wild-type (OreR) flies. Means ± SD. (E) Tyrosine metabolism pathway. *p<0.05, **p<0.01, ***p<0.001.
Figure 1—figure supplement 1.
Figure 1—figure supplement 1.. Age-dependent increase of GFP-tagged CG1461.
Immunoblot analysis of GFP and tubulin in 1-week, 4-week, and 8-week CG1461(TyrAm)-GFP-tagged flies.
Figure 2.
Figure 2.. CG1461 functions as Tyrosine Aminotransferase/TAT and is necessary to degrade tyrosine.
(A) Relative mRNA levels of CG1461 in tubulin-Gal80ts, tubulin-Gal4 flies expressing either no RNAi, control RNAi or three different CG1461 RNAi for 10 days. Means ± SD. *p<0.05, ***p<0.001 (B) Relative mRNA levels of CG1461 in wild-type and CG1461-deficient flies (both backcrossed to wild-type OreR flies). Means ± SD. ***p<0.001 (C) Feeding adult flies with 5 g/L of tyrosine significantly suppresses lifespan of flies with ubiquitous adult-onset downregulation of CG1461. Arrow indicates the beginning of tyrosine feeding. p<0.001. (D) Feeding adult flies with 5 g/L of tyrosine significantly suppresses lifespan of CG1461 mutant but not wild-type or heterozygous flies. Arrow indicates the beginning of tyrosine feeding. p<0.001. Box plots of relative levels of tyrosine (E) and phenylalanine (F) in wild-type and CG1461-deficient flies fed either control or high level of tyrosine (5 g/L) diet.
Figure 2—figure supplement 1.
Figure 2—figure supplement 1.. Tyrosine supplementation increases lifespan of wild-type OreR flies and increases expression of GFP-tagged CG1461.
Lifespan of male (A) and female (B) wild-type and CG1461-deficient flies (both backcrossed to wild-type OreR flies) fed with the range of tyrosine concentrations (1X = 0.5 g/L). Arrow indicates the beginning of tyrosine feeding. (C) Immunoblot analysis of GFP and tubulin in CG1461(TyrAm)-GFP-tagged flies fed either control or high-tyrosine diet. (D) Relative mRNA levels of CG1461, CG11796, Hgo, Faa, Hn, Rp49 in male and female flies. Means ± SD. Means ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 3.
Figure 3.. Whole-body and neuronal-specific downregulation of CG1461/Tyrosine Aminotransferase extends lifespan.
(A) Ubiquitous adult-onset expression of CG1461 RNAi-1 increases lifespan in females. p<0.0001. (B) Ubiquitous adult-onset expression of CG1461 RNAi-3 increases lifespan in females. p<0.0001. (C) Ubiquitous adult-onset expression of CG11796 RNAi increases lifespan in females. p<0.0001. (D) Ubiquitous adult-onset expression of Hgo RNAi increases lifespan in females. p<0.0001. (E) Fat body-specific adult-onset expression of CG1461 RNAi does not affect lifespan in females. (F) Intestine-specific adult-onset expression of CG1461 RNAi does not affect lifespan in females. (G) Neuronal-specific adult-onset expression of CG1461 RNAi-1 increases lifespan in females. p<0.0001. (H) Neuronal-specific adult-onset expression of CG1461 RNAi-2 increases lifespan in females. p<0.0001. (I) Neuronal-specific adult-onset expression of CG1461 RNAi-1, -2, and -3 increases lifespan in females. p<0.0001.
Figure 3—figure supplement 1.
Figure 3—figure supplement 1.. Neuronal-specific downregulation of Tyrosine Aminotransferase/TAT leads to metabolic reprogramming similar to the whole-body downregulation of TAT.
Box plots of relative levels of tyrosine (A), glucosamine (B), methylcysteine (C), and NADH (D) in elav-Gal4,tubulin-Gal80ts flies expressing either control RNAi or TAT RNAi. FDR adjusted p-value. *p<0.05, **p<0.01.
Figure 4.
Figure 4.. Downregulation of CG1461/Tyrosine Aminotransferase leads to reprogramming of metabolism related to mitochondrial function.
(A) Principal component analysis of tubulin-Gal4, tubulin-Gal80ts flies expressing either control RNAi or two different TAT RNAi. (B) Heat map showing the significantly and commonly changed metabolites in flies expressing two different TAT RNAi. Box plots of relative levels of tyrosine (C), lactate (D), glucosamine (E), nicotinamide (F), methylcysteine (G) in tubulin-Gal4,tubulin-Gal80ts flies expressing either control RNAi or two different TAT RNAi. (H) Metabolic Set Enrichment Analysis of the metabolites that changed significantly and commonly in flies expressing two different TAT RNAi. Box plots of relative levels of NADH (I), thiamine pyrophosphate (J), NADP (K) in tubulin-Gal4,tubulin-Gal80ts flies expressing either control RNAi or two different TAT RNAi.
Figure 4—figure supplement 1.
Figure 4—figure supplement 1.. Neuronal-specific downregulation of Tyrosine Aminotransferase/TAT increases levels of tyrosine-derived neurotransmitters.
Head levels of Histamine (A) and GABA (B) in CG1461/TAT wild-type (wt), heterozygous (het), and mutant (mut) flies. Head levels of DOPA (C), Octopamine (D), Dopamine (E), Tyramine (F) in elav-Gal4, tubulin-Gal80ts > control RNAi or TAT RNAi flies maintained either on the regular food or the food containing 5 mM of tyrosine for 2 days. Means ± SD. *p<0.05, **p<0.01, ***p<0.001. Feeding adult male (G) and female (H) OreR flies with 5 mM Octopamine, L-DOPA, or Tyramine starting day 14.
Figure 5.
Figure 5.. Whole-body downregulation of Tyrosine Aminotransferase/TAT elevates levels of tyrosine-derived neurotransmitters in fly heads.
Head levels of Tyrosine (A), DOPA (B), Dopamine (C), Tyramine (D), and Octopamine (E) in CG1461/TAT wild-type (wt), heterozygous (het), and mutant (mut) flies. Means ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 5—figure supplement 1.
Figure 5—figure supplement 1.. Suppression of complex I of ETC upregulates mRNA levels of enzymes in the tyrosine degradation pathway and decreases lifespan that can be partially rescued by supplementation of tyrosine.
(A) Relative mRNA levels of Faa, CG11796, Hgo in tubulin-Gal80ts, tubulin-Gal4 flies expressing either no RNAi, control RNAi or RNAi against different subunits of mitochondrial ETC Complex I – CG9762, NP15.6, mtacp1 for 10 days. Means ± SD. (B) Relative mRNA levels of CG1461/TAT, CG11796, Hgo, Faa, and Hn in tubulin-Gal80ts, elav-Gal4 flies expressing either control RNAi or RNAi against NP15.6. Lifespans of male (C) and female (D) tubulin-Gal80ts, tubulin-Gal4 flies expressing either control RNAi or RNAi against NP15.6 fed with the range of tyrosine concentrations (1X = 0.5 g/L). Neuronal counts (E) and histology (F) of hematoxylin-stained heads from flies with or without expression of wild-type human α-synuclein with or without three different RNAi against TAT. Means ± SD. *p<0.05, **p<0.01, ***p<0.001.
Figure 6.
Figure 6.. Mitochondrial dysfunction/neurodegeneration upregulates the level of CG1461/tyrosine aminotransferase.
(A) Relative mRNA levels of CG1461/TAT in Gal80ts; tubulin-Gal4 flies expressing either no RNAi, control RNAi or RNAi against different subunits of mitochondrial ETC – CG9762, SDHC, CG18809, CG5548, NP15.6, CG8680, CG1970, CG3214, mtacp1 for 10 days. Means ± SD. (B) Relative mRNA levels of CG1461/TAT and α -synuclein in heads of flies with or without expression of wild-type human α-synuclein. Means ± SD. (C) Relative mRNA levels of CG1461/TAT in tubulin-Gal80ts, tubulin-Gal4 flies overexpressing control or p60 (inhibitory subunit of InR), dTsc1/dTsc2 (TSC complex, dTOR inhibitor) or PGC1a/Spargel. Means ± SD. (D) Immunoblot analysis of GFP and tubulin in heads of 1-week and 3-week-old flies expressing either control or TAT RNAi under pan-neuronal driver (ElavGal4) in the presence of GstD-GFP. (E) Ubiquitous adult-onset downregulation of TAT prolongs lifespan under oxidative stress (10 mM Paraquat). (F) Relative mRNA levels of CG1461/TAT in tubulin-Gal80ts, tubulin-Gal4 flies expressing either control or NP15.6 RNAi and fed with 10 mM reduced Glutathione, 100 µM mitoQ, 10 mM methyl pyruvate, 100 µM Ibedenone, or 100 µM Tigecycline. Means ± SD (G) Relative mRNA levels of CG1461/TAT, CG11796, faa, and Hgo in tubulin-Gal80ts, tubulin-Gal4 flies expressing either control or NP15.6 RNAi and fed with either control or 100 µM Tigecycline. (H) Working model. *p<0.05, **p<0.01, ***p<0.001.
Figure 6—figure supplement 1.
Figure 6—figure supplement 1.. The effect of downregulation of tyrosine aminotransferase/TAT on GFP-tagged reporters relevant to aging.
(A) Schematic diagram of mitophagy/mitochondrial biogenesis regulation by Insulin signaling, TOR, and PGC1a/Spargel. (B) Immunoblot analysis of GFP and tubulin in heads of 1-week and 3-week-old tubulin-Gal80ts, elav-Gal4 flies in the presence of GFP-CL1 (B), hsp22-GFP (C), 10XSTAT92-GFP (D), Drs-GFP (E) reporters expressing either control or TAT RNAi. (F) Relative mRNA levels of CLPX, HSP10, HSP60, and HSP22 in tubulin-Gal80ts, tubulin-Gal4 flies expressing either control RNAi or NP15.6 RNAi and fed with either control or 100 µM Tigecycline. Means ± SD.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Area-Gomez E, Guardia-Laguarta C, Schon EA, Przedborski S. Mitochondria, OxPhos, and neurodegeneration: cells are not just running out of gas. Journal of Clinical Investigation. 2019;129:34–45. doi: 10.1172/JCI120848. - DOI - PMC - PubMed
    1. Avanesov AS, Ma S, Pierce KA, Yim SH, Lee BC, Clish CB, Gladyshev VN. Age- and diet-associated metabolome remodeling characterizes the aging process driven by damage accumulation. eLife. 2014;3:e02077. doi: 10.7554/eLife.02077. - DOI - PMC - PubMed
    1. Bach EA, Ekas LA, Ayala-Camargo A, Flaherty MS, Lee H, Perrimon N, Baeg GH. GFP reporters detect the activation of the Drosophila JAK/STAT pathway in vivo. Gene Expression Patterns. 2007;7:323–331. doi: 10.1016/j.modgep.2006.08.003. - DOI - PubMed
    1. Bahadorani S, Cho J, Lo T, Contreras H, Lawal HO, Krantz DE, Bradley TJ, Walker DW. Neuronal expression of a single-subunit yeast NADH-ubiquinone oxidoreductase (Ndi1) extends Drosophila lifespan. Aging Cell. 2010;9:191–202. doi: 10.1111/j.1474-9726.2010.00546.x. - DOI - PMC - PubMed
    1. Bai H, Kang P, Tatar M. Drosophila insulin-like peptide-6 (dilp6) expression from fat body extends lifespan and represses secretion of Drosophila insulin-like peptide-2 from the brain. Aging Cell. 2012;11:978–985. doi: 10.1111/acel.12000. - DOI - PMC - PubMed

Publication types

MeSH terms