Physiological Adaptations to Sugar Intake: New Paradigms from Drosophila melanogaster

Sugars are important energy sources, but high sugar intake poses a metabolic challenge and leads to diseases. Drosophila melanogaster is a generalist fruit breeder that encounters high levels of dietary sugars in its natural habitat. Consequently, Drosophila displays adaptive responses to dietary sugars, including highly conserved and unique metabolic adaptations not described in mammals. Carbohydrate homeostasis is maintained by a network comprising intracellular energy sensors, transcriptional regulators, and hormonal and neuronal mechanisms that together coordinate animal behavior, gut function, and metabolic flux. Here we give an overview of the physiological responses associated with sugar intake and discuss some of the emerging themes and applications of the Drosophila model in understanding sugar sensing and carbohydrate metabolism.

Redefining the metabolic landscape through TGF-beta/Activin signaling in Drosophila melanogaster

Wen Bin Chng

Multicellular organisms assess their nutritional state and adapt their metabolic landscape to match their energy demands. In this thesis, we address a longstanding conundrum of how an organism coordinates digestive enzyme expression in response to its nutritional status. Hydrolysis of food materials by endogenous or microbial-derived digestive enzymes precedes absorption through the gut lumen. Thus, modulation of endogenous digestive mechanism represents a pragmatic step to limit the uptake of nutrients when nutrients are in abundance. In Drosophila melanogaster, it is known that dietary glucose represses amylase transcript and enzymatic activity, in a process known as glucose repression. Here, we show that glucose reduces the expression of many carbohydrases, lipases, and glucose transporters, presumably to prevent nutritional overload after feeding. We found that the repression of carbohydrases is effected by nutritious sugars through the induction of the TGF-beta ligand, Dawdle. Interestingly, glucose repression of digestive enzymes is dependent upon the induction of Dawdle in the fat body, a tissue functionally analogous to the mammalian liver and adipose tissue. Dawdle is then transported via the circulation to activate TGFÎ²/Activin signaling in the midgut, leading to the repression of digestive enzymes expression. We later discovered that Dawdle expression in the fat body is regulated by the evolutionarily conserved Mio-bigmax transcription factor complex. In addition, expression of Dawdle must be tightly coordinated to reflect the nutritional state of adult flies, as flies overexpressing Dawdle are sensitive to starvation and have their TAG and glycogen stores rapidly depleted upon nutrient limitation. Thus, our results are consistent with a model whereby fat body derived Daw functions as a nutritional cue for carbohydrates to modulate metabolic responses in distal tissues. Its induction provides the nutritional signal to repress carbohydrate digestive capacity and stimulate peripheral insulin signaling. In conclusion, our study identifies the TGFÎ²/Activin signaling as a critical mechanism for carbohydrate homeostasis. Our results indicate that the fat body functions as a carbohydrate sensing tissue, which through the secretion of the TGFÎ² ligand, Daw, regulates organism-wide carbohydrate metabolism. Our work in Drosophila is likely to have a broader scientific impact by placing a pathway known for its role in development, immunity, and cancer, as an integral component for sugar sensing and metabolism.

EPFL2015

Genome Sequence of the Drosophila melanogaster Male-Killing Spiroplasma Strain MSRO Endosymbiont

Jeremy Keith Herren, Bruno Lemaitre, Ray Marcel Marin Florez, Juan Camilo Paredes Escobar

Spiroplasmas are helical and motile members of a cell wall-less eubacterial group called Mollicutes. Although all spiroplasmas are associated with arthropods, they exhibit great diversity with respect to both their modes of transmission and their effects on their hosts; ranging from horizontally transmitted pathogens and commensals to endosymbionts that are transmitted transovarially (i.e., from mother to offspring). Here we provide the first genome sequence, along with proteomic validation, of an endosymbiotic inherited Spiroplasma bacterium, the Spiroplasma poulsonii MSRO strain harbored by Drosophila melanogaster. Comparison of the genome content of S. poulsonii with that of horizontally transmitted spiroplasmas indicates that S. poulsonii has lost many metabolic pathways and transporters, demonstrating a high level of interdependence with its insect host. Consistent with genome analysis, experimental studies showed that S. poulsonii metabolizes glucose but not trehalose. Notably, trehalose is more abundant than glucose in Drosophila hemolymph, and the inability to metabolize trehalose may prevent S. poulsonii from overproliferating. Our study identifies putative virulence genes, notably, those for a chitinase, the H2O2-producing glycerol-3-phosphate oxidase, and enzymes involved in the synthesis of the eukaryote-toxic lipid cardiolipin. S. poulsonii also expresses on the cell membrane one functional adhesion-related protein and two divergent spiralin proteins that have been implicated in insect cell invasion in other spiroplasmas. These lipoproteins may be involved in the colonization of the Drosophila germ line, ensuring S. poulsonii vertical transmission. The S. poulsonii genome is a valuable resource to explore the mechanisms of male killing and symbiont-mediated protection, two cardinal features of many facultative endosymbionts. IMPORTANCE Most insect species, including important disease vectors and crop pests, harbor vertically transmitted endosymbiotic bacteria. These endosymbionts play key roles in their hosts' fitness, including protecting them against natural enemies and manipulating their reproduction in ways that increase the frequency of symbiont infection. Little is known about the molecular mechanisms that underlie these processes. Here, we provide the first genome draft of a vertically transmitted male-killing Spiroplasma bacterium, the S. poulsonii MSRO strain harbored by D. melanogaster. Analysis of the S. poulsonii genome was complemented by proteomics and ex vivo metabolic experiments. Our results indicate that S. poulsonii has reduced metabolic capabilities and expresses divergent membrane lipoproteins and potential virulence factors that likely participate in Spiroplasma-host interactions. This work fills a gap in our knowledge of insect endosymbionts and provides tools with which to decipher the interaction between Spiroplasma bacteria and their well-characterized host D. melanogaster, which is emerging as a model of endosymbiosis.

American Society for Microbiology2015

Elucidating regulatory mechanisms shaping the transcriptome and proteome of the gut immune response in Drosophila melanogaster

Michael Vincent Frochaux

Work on Drosophila melanogaster paved the way to our current understanding of modern genetics. Since then, this model organism has contributed greatly to various fields such as neurobiology, development, and immunology. The discovery and analysis of the various pathways of the Drosophila immune response led the way to the in-depth characterization of the gut immune response. Furthermore, a genome-wide association study on a population of Drosophila identified resistance to infection by the entomopathogenic bacteria Pseudomonas entomophila (P.e.) as a complex phenotype, with highly resistant and susceptible lines, and highlighted genes and pathways mediating inter-individual differences. However, the molecular mechanisms mediating the resistance to enteric infection remain largely uncharacterized. This study uses the same Drosophila population to analyze what are the molecular mechanisms mediating the resistance to enteric infection. We analyzed the genetic determinants of gene expression variation among the most resistant and susceptible lines in response to infection using expression quantitative trait loci (eQTL) analysis. We also characterized the mode of action of these eQTLs into cis- and trans-acting. Furthermore, we identified the gene nutcracker (ntc) as the only differentially-expressed gene between resistance classes. We then demonstrate that loss of function ntc mutants are significantly more susceptible to infection and then identify a single nucleotide polymorphism (SNP) located in a transcription factor binding site (TFBS) which affects the binding affinity of the transcription factor Broad leading to allele-specific expression variation of the gene ntc. If the transcriptomic response to infection has been thoroughly characterized, it is not the case for the proteomic response. We thus sought to characterize for the first time the proteomic response of the Drosophila to enteric infection. We performed mass spectrometry and RNA sequencing on dissected guts from flies infected by two Gram-negative bacteria, Erwinia carotovora carotovora 15 (Ecc15) or Pseudomonas entomophila, 4h and 16h after infection. We found that a large portion of the measurable proteome (12%) varies after infection and that protein changes are strongly time- and infection-dependent. We showed the relatively poor correlation between gene expression and protein abundance in the Drosophila enteric immune response. Finally, we performed a screen of several proteins that were identified in our proteomics work but had previously not been found when assessing the gene expression response to infection using loss-of-function mutants and identifying 7 genes that modulate overall susceptibility to P.e. infection. In summary, this work analyze the genetic determinants of gene expression variation in the DGRP population upon infection and characterize them according to their mode of action. Furthermore, we describe how a non-coding variant lowers resistance to infection by modulating ntc gene expression through altered Broad repressor binding. Finally, it provides the first comprehensive characterization of the Drosophila gut proteome upon infection by Gram-negative bacteria which can be the basis of future proteomic analysis of the enteric immune response

EPFL2019