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- W2912165268 abstract "Continuous oscillation of environmental conditions has led to the ubiquitous evolution of anticipatory mechanisms known as circadian rhythms. From the cellular to the organism level, these rhythms coordinate a plethora of physiological and behavioral responses. They provide organisms with elevated survival fitness, evident by the increased survival and reproductive rates of organisms maintaining clocks vs. those without a mechanism to time daily rhythms. Circadian rhythms are maintained by a conserved molecular clock orchestrated by a transcriptional/translational negative feedback loop (TTFL). Many of the proteins that organize this feedback loop are Intrinsically Disordered Proteins (IDPs), which lack a fixed or ordered three-dimensional structure. This includes the conserved disorder of the clock proteins in fungi, plants, and animals. Little is known about the impact of intrinsic disorder on clock proteins and this lack of comprehension is compounded by the fact that efficient techniques to understand the inherent nature of IDPs are only now developing. To address this gap in knowledge we developed a novel protocol to track the conformational dynamics of a core clock protein (FREQUENCY) in a fundamental clock model organism (Neurospora crassa). FREQUENCY (FRQ) is at the heart of the repressing arm of the TTFL and drives the length of the circadian period in N. crassa. Our protocol, circadian native fast parallel proteolysis (CRAFTY), utilizes a parallel proteolysis method in native conditions to determine the conformational shifts in FRQ over circadian time. CRAFTY yields biologically relevant information, enhancing our understanding of the importance of disorder in the circadian clock." @default.
- W2912165268 created "2019-02-21" @default.
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- W2912165268 date "2019-02-01" @default.
- W2912165268 modified "2023-10-18" @default.
- W2912165268 title "Characterizing Time-of-Day Conformational Changes in the IDP Frequency at the Heart of the Circadian Clock in N. Crassa using the Crafty Protocol" @default.
- W2912165268 doi "https://doi.org/10.1016/j.bpj.2018.11.1096" @default.
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