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Shekar, S.

Publications and source records attributed to Shekar, S..

2 recordsLinked to original sources

Fat body CLOCK restrains innate immunity and maintains survival under dietary stress in Drosophila

Both aging and a high-fat diet (HFD) dampen circadian gene transcription rhythms and promote chronic inflammation. How aging and HFD interact to affect peripheral molecular clocks and circadian behavior remains unclear. Using Drosophila melanogaster, we showed that aging and HFD additively dampened circadian behavior, with their adverse effects converging on the fat body (FB), a tissue that regulates systemic metabolism and innate immunity. Applying longitudinal in vivo bioluminescence recording in small, genetically defined cell populations, we found that molecular clocks in the FB were uniquely vulnerable to aging- and HFD-induced dampening of rhythm amplitude, whereas those in the clock neurons declined with age but were resistant to dietary stress. To test the consequences of this FB clock decline, we disrupted individual components of the core molecular clock specifically in the FB. We found that only CLOCK (CLK) disruption shortened lifespan on HFD, whereas disrupting its binding partner CYCLE (CYC), or the repressors PERIOD and TIMELESS, did not. Furthermore, CLK, but not CYC, disruption upregulated antimicrobial peptide expression in the FB, dampened behavioral rhythms, and suppressed locomotor activity, even though both CLK and CYC disruption comparably dampened clock gene oscillation in the FB. Together, these results indicate that FB CLK has a unique role in suppressing pro-inflammatory signals independently of CYC. Our findings provide insight into how stressors such as aging and HFD selectively disrupt the peripheral metabolic clock, and into the distinct roles of individual clock components, with implications for age-related inflammation and metabolic disease.

molecular biology↗

Inhibition of Proteasome Activity Facilitates Definitive Endodermal Specification of pluripotent Stem Cells by influencing YAP signaling

Understanding the molecular players that control the specification of definitive endoderm is imperative to obtain the homogenous population of pancreatic {beta}-cells from stem cells. Though the Ubiquitin proteasome system (UPS) has been envisaged as a crucial intracellular protein degradation system, its role in germ layer specification remains elusive. In this study, using a mouse embryonic stem cells model system (mESCs) we observed decreased proteasomal activity specifically during endoderm, but not in meso- or ecto-derm differentiation. Extraneous inhibition of proteasomal activity during differentiation enhanced the expression of endodermal genes specifically. Enhancing proteasomal activity by including the activator IU1 in the induction culture, inhibited definitive endodermal differentiation. Further, inhibiting proteasomal activity at the definitive endodermal stage resulted in enhanced generation of insulin-positive cells. A similar increase in endodermal gene expression by inhibiting proteasomal activity was observed in miPSC and hiPSC differentiated towards endodermal lineage. Mechanistic insight showed no contribution of endoplasmic reticulum unfolded protein response but revealed the involvement of the YAP signaling pathway in proteasome-inhibited enhanced endodermal differentiation. Unravelling the specific involvement of UPS in endodermal cell fate specification in pluripotent stem cells paves the way for obtaining better qualitative and quantitative definitive endodermal cells for plausible cellular therapy in the future.

cell biology↗