Chromatin organization plays a crucial role in gene regulation by controlling the accessibility of DNA to transcription machinery. While significant progress has been made understanding regulatory clock proteins circadian rhythms, how chromatin affects rhythms remains poorly understood. Here, we employed ATAC-seq (Assay for Transposase-Accessible with Sequencing) on FAC-sorted Drosophila neurons assess genome-wide at dawn and dusk over cycle. We observed oscillations promoter enhancer...

Circadian clocks regulate ∼24-h oscillations in gene expression, behavior, and physiology. While the genetic molecular mechanisms of circadian rhythms are well characterized, what remains poorly understood intracellular dynamics clock components how they affect rhythms. Here, we elucidate spatiotemporal organization core proteins genes Drosophila neurons. Using high-resolution imaging DNA-fluorescence situ hybridization techniques, demonstrate that (PERIOD CLOCK) organized into a few...

Adult stem cells continuously supply differentiated throughout the life of organisms. This increases risk replicative senescence or neoplastic transformation due to mutations that accumulate over many rounds DNA replication. The immortal strand hypothesis proposes reduce accumulation replication-induced by retaining older template strands. Other models have also been proposed in which asymmetrically segregate chromosome strands for other reasons, such as retention epigenetic memories....

Abstract Calorimetry has been widely used in metabolic studies, but direct measurements from individual small biological model organisms such as C. elegans or isolated single cells have limited by poor sensitivity of existing techniques and difficulties resolving very heat outputs. Here, careful thermal engineering, we developed a robust, highly sensitive bio-compatible calorimetric platform that features resolution ~270 pW—more than 500-fold improvement over the most calorimeter previously...

Adult stem cells continuously supply differentiated throughout the life of organisms. This increases risk replicative senescence or neoplastic transformation due to mutations that accumulate over many rounds DNA replication. The immortal strand hypothesis proposes reduce accumulation replication-induced by retaining older template strands. Other models have also been proposed in which asymmetrically segregate chromosome strands for other reasons, such as retention epigenetic memories....

Abstract Significant recent evidence suggests that metabolism is intricately linked to the regulation and dysfunction of complex cellular physiological responses ranging from altered metabolic programs in cancers aging circadian rhythms molecular clocks. While pathways their fundamental control mechanisms are well established, precise underpinning, for example, enzymatic pathway control, substrate preferences or rates, remain far less certain. Comprehensive, continuous studies on model...

Circadian clocks are ∼24-h timekeepers that control rhythms in almost all aspects of our behavior and physiology. While it is well known subcellular localization core clock proteins plays a critical role circadian regulation, very little about the spatiotemporal organization mRNAs its generating rhythms. Here we describe streamlined single molecule Fluorescence In Situ Hybridization (smFISH) protocol fully automated analysis pipeline to precisely quantify number location Clock ,...

We thank the authors of correspondence ‘Overlooked areas for attention needed sound evaluation DNA strand inheritance patterns in Drosophila male germline stem cells’ raising important points related to our article ‘Drosophila cells do not asymmetrically segregate chromosome strands’ (Yadlapalli et al., 2011). welcome opportunity extend discussion and explain more fully thoroughly. Note that author(s) this initially did wish reveal their identity until reply are published. Therefore, we have...

The study of protein subcellular localization, dynamics, and regulation in live cells has been profoundly transformed by the advent techniques that allow tagging endogenous genes to produce fluorescent fusion proteins. These methods enable researchers visualize behavior real time, providing valuable insights into their functions interactions within cellular environment. Many current gene studies employ a two-step process where visible markers, such as eye color changes, are used identify...

Achieving micro-kelvin (µK) temperature stability is critical for many calorimetric applications. For example, sub-nanowatt resolution biocalorimetry requires stabilization of the calorimeter to µK levels. Here, we describe how can be accomplished in a prototypical system consisting two nested shields and suspended capillary tube, which well suited Specifically, show that by employing with µTorr-levels vacuum space between them as precise feedback control (performed using high-resolution...

ABSTRACT The ability to tag endogenous genes create fluorescent fusion proteins has revolutionized the studies of protein subcellular localization and dynamics regulation in live cells. Here, we report a precise, rapid, one-step gene tagging system Drosophila melanogaster , which allows us screen for engineered lines without visible eye marker. Specifically, developed an efficient screening strategy identification homologous integration events by employing PCR-based method characterize...

Abstract Chromatin organization plays a crucial role in gene regulation by controlling the accessibility of DNA to transcription machinery. While significant progress has been made understanding regulatory clock proteins circadian rhythms, how chromatin affects rhythms remains poorly understood. Here, we employed ATAC-seq (Assay for Transposase-Accessible with Sequencing) on FAC-sorted Drosophila neurons assess genome-wide over cycle. We observed oscillations at promoter and enhancer regions...

ABSTRACT Circadian clocks regulate ∼24 hour oscillations in gene expression, behavior, and physiology. While the molecular neural mechanisms of circadian rhythms are well characterized, how cellular organization clock components controls regulation remains poorly understood. Here, we elucidate proteins by controlling spatiotemporal genes. Using high-resolution live imaging techniques demonstrate that Drosophila concentrated a few discrete foci organized at nuclear envelope; these results...