Total Solid-Phase Synthesis of Biologically Active Drosophila Insulin-Like Peptide 2 (DILP2)

Lin, Feng; Hossain, Mohammed Akhter; Post, Stephanie; Karashchuk, Galina; Tatar, Marc; Meyts, Pierre De; Wade, John D.

doi:10.1071/ch16626

Cited by 12 publications

(12 citation statements)

References 31 publications

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“…Drosophila insulin-like peptide 2 and DILP5 are similarly expressed and released from brain medial secretory neurons in adult Drosophila , yet appear to have distinct roles in glucose and protein metabolism and in aging ( 8 , 16 ). To understand how these related ligands generate different outcomes in vivo , we treated Drosophila S2 cells in culture with purified synthetic DILP2 ( 37 ) and recombinant DILP5 ( 36 ). At doses ranging from 0.1 to 100 nM, DILP2 and DILP5 stimulated comparable increases in Akt phosphorylation at Ser505 (Figure 2 A), suggesting they have similar potency.…”

Section: Resultsmentioning

confidence: 99%

“…For overnight serum depletion, cells were held for 16 h in Schneider’s media without FBS, supplemented with 0.5% BSA and 25 mM HEPES. DILP5 peptide ( 36 ) stock at 50 μM and DILP2 peptide stock ( 37 ) at 100 μM stimulated overnight depleted cells compared with control depleted cells treated in parallel with an equal volume of 0.001 N HCl solvent (vehicle). The dose and duration of DILP stimulation for each experiment is detailed in the figure legend.…”

Section: Methodsmentioning

confidence: 99%

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Drosophila Insulin-Like Peptides DILP2 and DILP5 Differentially Stimulate Cell Signaling and Glycogen Phosphorylase to Regulate Longevity

et al. 2018

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Insulin and IGF signaling (IIS) is a complex system that controls diverse processes including growth, development, metabolism, stress responses, and aging. Drosophila melanogaster IIS is propagated by eight Drosophila insulin-like peptides (DILPs), homologs of both mammalian insulin and IGFs, with various spatiotemporal expression patterns and functions. DILPs 1–7 are thought to act through a single Drosophila insulin/IGF receptor, InR, but it is unclear how the DILPs thereby mediate a range of physiological phenotypes. We determined the distinct cell signaling effects of DILP2 and DILP5 stimulation upon Drosophila S2 cells. DILP2 and DILP5 induced similar transcriptional patterns but differed in signal transduction kinetics. DILP5 induced sustained phosphorylation of Akt, while DILP2 produced acute, transient Akt phosphorylation. Accordingly, we used phosphoproteomic analysis to identify distinct patterns of non-genomic signaling induced by DILP2 and DILP5. Across all treatments and replicates, 5,250 unique phosphopeptides were identified, representing 1,575 proteins. Among these peptides, DILP2, but not DILP5, dephosphorylated Ser15 on glycogen phosphorylase (GlyP), and DILP2, but not DILP5, was subsequently shown to repress enzymatic GlyP activity in S2 cells. The functional consequences of this difference were evaluated in adult Drosophila dilp mutants: dilp2 null adults have elevated GlyP enzymatic activity relative to wild type, while dilp5 mutants have reduced GlyP activity. In flies with intact insulin genes, GlyP overexpression extended lifespan in a Ser15 phosphorylation-dependent manner. In dilp2 mutants, that are otherwise long-lived, longevity was repressed by expression of phosphonull GlyP that is enzymatically inactive. Overall, DILP2, unlike DILP5, signals to affect longevity in part through its control of phosphorylation to deactivate glycogen phosphorylase, a central modulator of glycogen storage and gluconeogenesis.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Drosophila Insulin-Like Peptides DILP2 and DILP5 Differentially Stimulate Cell Signaling and Glycogen Phosphorylase to Regulate Longevity

et al. 2018

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“…DILP1 through DILP5 are most closely related to vertebrate insulin, whereas DILP6 is the only IGF-like peptide in Drosophila ( Okamoto et al 2009 ). These six DILPs are believed to act through the single insulin RTK InR ( Fernandez et al 1995 ; Chen et al 1996 ; Brogiolo et al 2001 ), although only DILP2 and DILP5 have been assayed biochemically for InR activity ( Sajid et al 2011 ; Lin et al 2017 ; Post et al 2018a ). DILP7 and DILP8 appear to be more closely related to human relaxin-family molecules than to insulin/IGF.…”

Section: Body-size Controlmentioning

confidence: 99%

Regulation of Body Size and Growth Control

Texada¹,

Kaburagi²,

Rewitz³

2020

Genetics

129

146

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The control of body and organ growth is essential for the development of adults with proper size and proportions, which is important for survival and reproduction. In animals, adult body size is determined by the rate and duration of juvenile growth, which are influenced by the environment. In nutrient-scarce environments in which more time is needed for growth, the juvenile growth period can be extended by delaying maturation, whereas juvenile development is rapidly completed in nutrient-rich conditions. This flexibility requires the integration of environmental cues with developmental signals that govern internal checkpoints to ensure that maturation does not begin until sufficient tissue growth has occurred to reach a proper adult size. The Target of Rapamycin (TOR) pathway is the primary cell-autonomous nutrient sensor, while circulating hormones such as steroids and insulin-like growth factors are the main systemic regulators of growth and maturation in animals. We discuss recent findings in Drosophila melanogaster showing that cell-autonomous environment and growth-sensing mechanisms, involving TOR and other growth-regulatory pathways, that converge on insulin and steroid relay centers are responsible for adjusting systemic growth, and development, in response to external and internal conditions. In addition to this, proper organ growth is also monitored and coordinated with whole-body growth and the timing of maturation through modulation of steroid signaling. This coordination involves interorgan communication mediated by Drosophila insulin-like peptide 8 in response to tissue growth status. Together, these multiple nutritional and developmental cues feed into neuroendocrine hubs controlling insulin and steroid signaling, serving as checkpoints at which developmental progression toward maturation can be delayed. This review focuses on these mechanisms by which external and internal conditions can modulate developmental growth and ensure proper adult body size, and highlights the conserved architecture of this system, which has made Drosophila a prime model for understanding the coordination of growth and maturation in animals.

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“…Therefore, a deficiency of insulin might be the reason for the decrease of INSRβ phosphorylation. However, eight ILPs were identified by BLAST searching of the H. armigera genome using the sequences of 38 Bombyx mori ILPs, 8 Drosophila ILPs ( 25 ), and 10 Homo sapiens ILPs ( Fig. S2 ), and which ILP binds to INSR was not known; therefore, the expression levels of all Ilp genes were detected to address the insulin levels.…”

Section: Resultsmentioning

confidence: 99%

The steroid hormone 20-hydroxyecdysone counteracts insulin signaling via insulin receptor dephosphorylation

Yao

Zhao

et al. 2021

Journal of Biological Chemistry

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The insulin receptor (INSR) binds insulin to promote body growth and maintain normal blood glucose levels. While it is known that steroid hormones such as estrogen and 20-hydroxyecdysone counteract insulin function, the molecular mechanisms responsible for this attenuation remain unclear. In the present study, using the agricultural pest lepidopteran Helicoverpa armigera as a model, we proposed that the steroid hormone 20-hydroxyecdysone (20E) induces dephosphorylation of INSR to counteract insulin function. We observed high expression and phosphorylation of INSR during larval feeding stages that decreased during metamorphosis. Insulin upregulated INSR expression and phosphorylation, whereas 20E repressed INSR expression and induced INSR dephosphorylation in vivo . Protein tyrosine phosphatase 1B (PTP1B, encoded by Ptpn1 ) dephosphorylated INSR in vivo . PTEN (phosphatase and tensin homolog deleted on chromosome 10) was critical for 20E-induced INSR dephosphorylation by maintaining the transcription factor Forkhead box O (FoxO) in the nucleus, where FoxO promoted Ptpn1 expression and repressed Insr expression. Knockdown of Ptpn1 using RNA interference maintained INSR phosphorylation, increased 20E production, and accelerated pupation. RNA interference of Insr in larvae repressed larval growth, decreased 20E production, delayed pupation, and accumulated hemolymph glucose levels. Taken together, these results suggest that a high 20E titer counteracts the insulin pathway by dephosphorylating INSR to stop larval growth and accumulate glucose in the hemolymph.

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Total Solid-Phase Synthesis of Biologically Active Drosophila Insulin-Like Peptide 2 (DILP2)

Cited by 12 publications

References 31 publications

Drosophila Insulin-Like Peptides DILP2 and DILP5 Differentially Stimulate Cell Signaling and Glycogen Phosphorylase to Regulate Longevity

Drosophila Insulin-Like Peptides DILP2 and DILP5 Differentially Stimulate Cell Signaling and Glycogen Phosphorylase to Regulate Longevity

Regulation of Body Size and Growth Control

The steroid hormone 20-hydroxyecdysone counteracts insulin signaling via insulin receptor dephosphorylation

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