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Gene Expression Analysis of Fat Body of Drosophila melanogaster Subjected to Sir2 Overexpression or Dietary Restriction

ABSTRACT: Sir2 is the most intensively discussed longevity gene in current aging research. Although the gene encoding for a NAD+-dependent histone deacetylase initially was found to extend lifespan of various organisms ranging from yeast to mammals, serious doubts regarding its role in longevity have been expressed recently. In this study, we tested whether tissue-specific overexpression of Sir2 in the adult fat body can extend lifespan when compared to genetically identical controls. We also wanted to elucidate the mechanisms by which fat body Sir2 promotes longevity by studying the phenotypic and transcriptional changes in the fat body. We found that moderate (3-fold) Sir2 overexpression in the fat body during adulthood only can promote longevity in both sexes by roughly 13 %. In addition, we obtained transcriptional profiles elicited by this overexpression and propose a role for Sir2 in lipid droplet biology especially under conditions of starvation. Furthermore, our data do not support the idea of Sir2 mediating the response to dietary restriction (DR) because transcriptional profiles of fat bodies after DR or Sir2 overexpression do not match. This study provides additional independent evidence for the concept of Sir2 as a longevity gene and as a promising pharmacological target to cure age-related diseases.

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE45974 | GEO | 2013/05/01

SECONDARY ACCESSION(S): PRJNA196804

REPOSITORIES: GEO

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Gene Expression Analysis of Fat Body of Drosophila melanogaster Subjected to Sir2 Overexpression or Dietary Restriction

Project description:Sir2 is the most intensively discussed longevity gene in current aging research. Although the gene encoding for a NAD+-dependent histone deacetylase initially was found to extend lifespan of various organisms ranging from yeast to mammals, serious doubts regarding its role in longevity have been expressed recently. In this study, we tested whether tissue-specific overexpression of Sir2 in the adult fat body can extend lifespan when compared to genetically identical controls. We also wanted to elucidate the mechanisms by which fat body Sir2 promotes longevity by studying the phenotypic and transcriptional changes in the fat body. We found that moderate (3-fold) Sir2 overexpression in the fat body during adulthood only can promote longevity in both sexes by roughly 13 %. In addition, we obtained transcriptional profiles elicited by this overexpression and propose a role for Sir2 in lipid droplet biology especially under conditions of starvation. Furthermore, our data do not support the idea of Sir2 mediating the response to dietary restriction (DR) because transcriptional profiles of fat bodies after DR or Sir2 overexpression do not match. This study provides additional independent evidence for the concept of Sir2 as a longevity gene and as a promising pharmacological target to cure age-related diseases. 6 groups of sample types were included in the experiment: a) females overexpressing Sir2 in the fat body b) female controls c) males overexpressing Sir2 in the fat body d) male controls e) wildtype females subjected to DR f) wildtype females feeding on a normal diet. 3 biological replicates were included per group.

2013-05-01 | E-GEOD-45974 | biostudies-arrayexpress

Expression Data From Dietary Restriction, p53 Knockdown and sir2 Overexpression Mutants

Project description:Expression data from four different lifespan-extending conditions: dietary restriction in two different genetic backgrounds (canton-s and a yw, w1118 combination), sir2 overexpression and p53 knockdown (+/-). Comparison of significantly over and under-expressed genes reveals a signature for dietary restriction and lifespan extension. Abstract A major challenge in translating the positive effects of dietary restriction (DR) for the improvement of human health is the development of therapeutic mimics. One approach to finding DR mimics is based upon identification of the proximal effectors of DR life span extension. Whole genome profiling of DR in Drosophila shows a large number of changes in gene expression, making it difficult to establish which changes are involved in life span determination as opposed to other unrelated physiological changes. We used comparative whole genome expression profiling to discover genes whose change in expression is shared between DR and two molecular genetic life span extending interventions related to DR, increased dSir2 and decreased Dmp53 activity. We find twenty-one genes shared among the three related life span extending interventions. One of these genes, takeout, thought to be involved in circadian rhythms, feeding behavior and juvenile hormone binding is also increased in four other life span extending conditions: Rpd3, Indy, chico and methuselah. We demonstrate takeout is involved in longevity determination by specifically increasing adult takeout expression and extending life span. These studies demonstrate the power of comparative whole genome transcriptional profiling for identifying specific downstream elements of the DR life span extending pathway. 42 samples were used in the analysis. Sir2 overexpression flies and the yw, w1118 background DR flies had the same control. Flies were collected at ages 10 and 40 days. Tissue is whole body females.

2011-01-20 | E-GEOD-26726 | biostudies-arrayexpress

Xbp1 overexpression in the gut and fat body extend Drosophila lifespan

Project description:Manipulating the activity of transcription factor (TF) to regulate animal’s longevity is achieved in different species. However, deciphering the pro-longevity transcriptional programme triggered by the TFs activity is challenging. One obstacle is the multifunctional feature of TFs. The physiological functions of single TF could be diverse and tissues-dependent. The other is the gene regulatory network among the TFs, where multiple TFs can have synergetic or antagonistic impact on the same target genes. Here, we show Xbp1s overexpression in gut and fat body can extend lifespan in Drosophila. Importantly, Xbp1s activity triggers distinct transcriptional programmes in the two tissues, and Xbp1s induction in both tissues are beneficial for longevity. Furthermore, we reveal a pro-longevity gene regulatory network in the fat body, where Xbp1s and dFOXO impinge on the same target genes and induce identical transcriptional outcomes. dfoxo overexpression requires Xbp1 to extend lifespan. Lastly, inducing Xbp1u, the precursor of Xbp1s, can also extend lifespan without triggering massive transcriptome change.

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Expression Data From Dietary Restriction, p53 Knockdown and sir2 Overexpression Mutants

2011-01-20 | GSE26726 | GEO

Proteomic analysis of dietary restriction in yeast reveals a role for Hsp26 in lifespan extension C4PR_LIV

Project description:Dietary restriction (DR) has been shown to increase lifespan in organisms ranging from yeast to mammals. This suggests that the underlying mechanisms may be evolutionarily conserved. Indeed, upstream signalling pathways, such as TOR, are strongly linked to DR-induced longevity in various organisms. However, the downstream effector proteins that ultimately mediate lifespan extension are less clear. To shed light on this, we used a proteomic approach on budding yeast. Our reasoning was that analysis of proteome-wide changes in response to DR might enable the identification of proteins that mediate its physiological effects, including lifespan extension. Of over 2500 proteins we identified by liquid chromatography-mass spectrometry, 183 were significantly altered in expression by at least 3-fold in response to DR. Most of these proteins were mitochondrial and/or had clear links to respiration and metabolism. Indeed, direct analysis of oxygen consumption confirmed that mitochondrial respiration was increased several-fold in response to DR. In addition, several key proteins involved in mating, including Ste2 and Ste6, were downregulated by DR. Consistent with this, shmoo formation in response to α-factor pheromone was reduced by DR, thus confirming the inhibitory effect of DR on yeast mating. Finally, we found that Hsp26, a member of the conserved small heat shock protein (sHSP) family, was upregulated by DR and that overexpression of Hsp26 extends yeast replicative lifespan. As overexpression of sHSPs in C. elegans and Drosophila has previously been shown to extend lifespan, our data on yeast Hsp26 suggest that sHSPs may be universally conserved effectors of longevity.

2022-02-17 | PXD025004 | Pride

Xbp1 overexpression in the gut and fat body extend Drosophila lifespan

Project description:Xbp1 overexpression in the gut and fat body extend Drosophila lifespan

| PRJNA1032092 | ENA

Sir2 mutants versus Controls at 2 weeks of age

Project description:Although SIRT1 plays a central role in maintaining metabolic homeostasis, the molecular mechanisms remain unclear. Here we show that loss of the Drosophila SIRT1 homolog sir2 leads to the progressive onset of diabetic phenotypes, similar to studies of SIRT1 in mice. Sir2 function is both necessary and sufficient in the fat body to maintain peripheral insulin sensitivity. This activity is mediated by the Drosophila HNF4 nuclear receptor, which is deacetylated and stabilized through protein interactions with Sir2. This study demonstrates that the key metabolic activities of SIRT1 have been conserved through evolution and establishes HNF4 as a critical downstream target. 4 sir2 mutant, 4 control samples, independent biological replicates

2016-02-29 | E-GEOD-72947 | biostudies-arrayexpress

Inhibition of S6K lowers age-related inflammation and immunosenescence and increases lifespan via the endolysosomal system

Project description:The nutrient-sensing Target of Rapamycin complex 1 (TORC1) is an evolutionarily conserved regulator of longevity. S6 kinase (S6K) is an essential downstream mediator for the effect of TORC1 on longevity. However, mechanistic insights on how TORC1-S6K signalling promotes lifespan and healthspan are still limited. Here we show that activity of S6K in the Drosophila fat body is essential for rapamycin-mediated longevity. Fat-body-specific activation of S6K blocked lifespan extension upon rapamycin feeding and induced accumulation of multilamellar lysosomal enlargements. Besides, fat body-specific S6K knockdown extended lifespan in files. We performed proteomics for Drosophila fat body to explore the fat body-specific regulation of protein expression by two separate datasets: fat body-specific S6K activation (Lsp2GS>S6KCA) with rapamycin treatment; and fat body-specific S6K inhibition (Lsp2GS>S6KRNAi). To assess if the age-prolonging mechanisms of TORC1-S6K signalling are conserved between flies and mammals, we assessed the impact of rapamycin treatment in the proteome of liver from C3B6F1 mice (F1 hybrids of C3H/HeOuJ females and C57Bl/6N males).

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Sir2 mutants versus Controls at 2 weeks of age

2016-02-29 | GSE72947 | GEO

Transcriptional profiling of fruit flies overexpressing the Atg1 gene

Project description:Overexpression of Atg1 using either weaker CGGAL4 or stronger HRGAL4 driver. Both drivers have same expression pattern and are specific for intestine, Malpighian tubules and fat body of the fruit fly Drosophila melanogaster. Weaker Atg1 overexpression results in longer lifespan whereas stronger Atg1 overexpression shortens lifespan compared to controls. Overexpressor lines were combined with tubGAL80ts, which is a temperature sensitive GAL4 repressor. This enabled induction of Atg1 overexpression in day-2 adult fly, thereby bypassing development and any potential developmental effects that Atg1 might have. Fly eggs of appropriate genotype were raised under standard density at 18C and then emerged flies were switched to 27C at day 2 of adulthood for Atg1 overexpression, and were kept at 27C onwards for longevity analyses. Dissected tissue was collected for the transcriptomic analysis at two weeks of age.

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