scholarly journals period-1 encodes an ATP-dependent RNA helicase that influences nutritional compensation of the Neurospora circadian clock

2015 ◽  
Vol 112 (51) ◽  
pp. 15707-15712 ◽  
Author(s):  
Jillian M. Emerson ◽  
Bradley M. Bartholomai ◽  
Carol S. Ringelberg ◽  
Scott E. Baker ◽  
Jennifer J. Loros ◽  
...  

Mutants in the period-1 (prd-1) gene, characterized by a recessive allele, display a reduced growth rate and period lengthening of the developmental cycle controlled by the circadian clock. We refined the genetic location of prd-1 and used whole genome sequencing to find the mutation defining it, confirming the identity of prd-1 by rescuing the mutant circadian phenotype via transformation. PRD-1 is an RNA helicase whose orthologs, DDX5 [DEAD (Asp-Glu-Ala-Asp) Box Helicase 5] and DDX17 in humans and DBP2 (Dead Box Protein 2) in yeast, are implicated in various processes, including transcriptional regulation, elongation, and termination, ribosome biogenesis, and mRNA decay. Although prd-1 mutants display a long period (∼25 h) circadian developmental cycle, they interestingly display a WT period when the core circadian oscillator is tracked using a frq-luciferase transcriptional fusion under conditions of limiting nutritional carbon; the core oscillator in the prd-1 mutant strain runs with a long period under glucose-sufficient conditions. Thus, PRD-1 clearly impacts the circadian oscillator and is not only part of a metabolic oscillator ancillary to the core clock. PRD-1 is an essential protein, and its expression is neither light-regulated nor clock-regulated. However, it is transiently induced by glucose; in the presence of sufficient glucose, PRD-1 is in the nucleus until glucose runs out, which elicits its disappearance from the nucleus. Because circadian period length is carbon concentration-dependent, prd-1 may be formally viewed as a clock mutant with defective nutritional compensation of circadian period length.

Genetics ◽  
1978 ◽  
Vol 88 (2) ◽  
pp. 255-265
Author(s):  
Jerry F Feldman ◽  
Cheryl A Atkinson

ABSTRACT A circadian clock mutant of Neurospora crassa with a period length of about 25.8 hours (4 hr longer than wild type) has been isolated after mutagenesis of the band strain. This mutant, called frq-5, segregates as a single nuclear gene, maps near the centromere on linkage group III, and is unlinked to four previously described clock mutants clustered on linkage group VII R (Feldman and Hoyle 1973, 1976). frq-5 differs from the other clock mutants in at least two other respects: (1) it is recessive in heterokaryons, and (2) it grows at about 60% the rate of the parent band strain on both minimal and complete media. Double mutants between frq-5 and each of the other clock mutants show additivity of period length-two long period mutants produce a double mutant whose period length is longer than either of the two single mutants, while a long and a short period double mutant has an intermediate period length. Although slow growth and long periodicity of frq-5 have segregated together among more than 300 progeny, slow growth per se is not responsible for the long period, since all the double mutants have the slow growth characteristic of frq-5, but have period lengths both shorter and longer than wild type.


2019 ◽  
Vol 34 (2) ◽  
pp. 218-223 ◽  
Author(s):  
Manpreet Kaur ◽  
Amy Ng ◽  
Pyonghwa Kim ◽  
Casey Diekman ◽  
Yong-Ick Kim

Cyanobacteria contain a circadian oscillator that can be reconstituted in vitro. In the reconstituted circadian oscillator, the phosphorylation state of KaiC oscillates with a circadian period, spending about 12 h in the phosphorylation phase and another 12 h in the dephosphorylation phase. Although some entrainment studies have been performed using the reconstituted oscillator, they were insufficient to fully explain entrainment mechanisms of the cyanobacterial circadian clock due to the lack of input pathway components in the in vitro oscillator reaction mixture. Here, we investigate how an input pathway component, CikA, affects the phosphorylation state of KaiC in vitro. In general, CikA affects the amplitude and period of the circadian oscillation of KaiC phosphorylation by competing with KaiA for the same binding site on KaiB. In the presence of CikA, KaiC switches from its dephosphorylation phase to its phosphorylation phase prematurely, due to an early release of KaiA from KaiB as a result of competitive binding between CikA and KaiA. This causes hyperphosphorylation of KaiC and lowers the amplitude of the circadian oscillation. The period of the KaiC phosphorylation oscillation is shortened by adding increased amounts of CikA. A constant period can be maintained as CikA is increased by proportionally decreasing the amount of KaiA. Our findings give insight into how to reconstitute the cyanobacterial circadian clock in vitro by the addition of an input pathway component, and explain how this affects circadian oscillations by directly interacting with the oscillator components.


2020 ◽  
Author(s):  
J. Romário F. de Melo ◽  
Annelie Gutsch ◽  
Joëlle De Caluwé ◽  
Jean-Christophe Leloup ◽  
Didier Gonze ◽  
...  

AbstractThe circadian clock coordinates the physiological response of a biological system to day and night rhythms through complex loops of transcriptional/ translational regulation. It can respond to external stimuli and adjust generated circadian oscillations accordingly to keep an endogenous period close to 24 h. To date, the interaction between nutritional status and circadian rhythms in plants is poorly understood. Magnesium (Mg) is essential for numerous biological processes in plants and its homeostasis is crucial to maintain optimal development and growth. Magnesium deficiency in young Arabidopsis thaliana seedlings increased the circadian period of pCCA1:LUC oscillations and dampened its amplitude in constant light in a dose-dependent manner. Although circadian period increase by Mg deficiency was light dependent, it did not depend on active photosynthesis. Mathematical modelling of the Mg input to the circadian clock reproduced the experimental increase of the circadian period and suggested that Mg is likely to affect global transcription/translation levels rather than a single component of the circadian oscillator. The model prediction was supported by a synergistic interaction between Mg deficiency and cyclohexamide, an inhibitor of translation. These findings suggest that proper Mg supply is required to support proper timekeeping in plants.One sentence summaryMagnesium maintains the circadian period in Arabidopsis seedlings and interferes with the circadian oscillator most likely through translational mechanisms.


2021 ◽  
Author(s):  
Jeffrey N. Schellinger ◽  
Qifei Sun ◽  
John M. Pleinis ◽  
Sung-Wan An ◽  
Jianrui Hu ◽  
...  

Central pacemaker neurons regulate circadian rhythms and undergo diurnal variation in electrical activity in mammals and flies. In mammals, circadian variation in the intracellular chloride concentration of pacemaker neurons has been proposed to influence the response to GABAergic neurotransmission through GABAA receptor chloride channels. However, results have been contradictory, and a recent study demonstrated circadian variation in pacemaker neuron chloride without an effect on GABA response. Therefore, whether and how intracellular chloride regulates circadian rhythms remains controversial. Here, we demonstrate a signaling role for intracellular chloride in the Drosophila ventral lateral (LNv) pacemaker neurons. In control flies, intracellular chloride increases in LNv neurons over the course of the morning. Chloride transport through the sodium-potassium-2-chloride (NKCC) and potassium-chloride (KCC) cotransporters is a major determinant of intracellular chloride concentrations. Drosophila melanogaster with loss-of-function mutations in the NKCC encoded by Ncc69 have abnormally low intracellular chloride six hours after lights on, and a lengthened circadian period. Loss of kcc, which is expected to increase intracellular chloride, suppresses the long-period phenotype of Ncc69 mutant flies. Activation of a chloride-inhibited kinase cascade, consisting of the WNK (With No Lysine (K)) kinase and its downstream substrate, Fray, is necessary and sufficient to prolong period length. Fray activation of an inwardly rectifying potassium channel, Irk1, is also required for the long-period phenotype. These results indicate that the NKCC-dependent rise in intracellular chloride in Drosophila LNv pacemaker neurons restrains WNK-Fray signaling and overactivation of an inwardly rectifying potassium channel to maintain normal circadian period length.


2018 ◽  
Author(s):  
JingJing Liu ◽  
Xianlin Zou ◽  
Tetsuya Gotoh ◽  
Anne M. Brown ◽  
Liang Jiang ◽  
...  

ABSTRACTThe circadian clock relies on post-translational modifications to set the timing for degradation of core regulatory components and, thus, sets clock progression. Ubiquitin-modifying enzymes targeting clock components for degradation are known to mostly recognize phosphorylated substrates. A case in point is the circadian factor PERIOD 2 (PER2) whose phospho-specific turnover involves its recognition by β-transducin repeat containing proteins (β-TrCPs). Yet, the existence of this unique mode of regulation of PER2’s stability falls short of explaining persistent oscillatory phenotypes reported in biological systems lacking functional elements of the phospho-dependent PER2 degradation machinery.In this study, we challenge the phosphorylation-centric view that PER2 degradation enhances circadian rhythm robustness by i) identifying the PER2:MDM2 endogenous complex, ii) establishing PER2 as a previously uncharacterized substrate for MDM2, iii) revealing an alternative phosphorylation-independent mechanism for PER2 ubiquitin-mediated degradation, iv) pinpointing residues for ubiquitin modification, and v) establishing the importance of MDM2-mediated PER2 turnover for defining the circadian period length. Our results not only expand MDM2’s suite of specific substrates beyond the cell cycle to include circadian components but also uncover novel regulatory players that likely impact our view of how other mechanisms crosstalk and modulate the clock itself.


2021 ◽  
Vol 12 ◽  
Author(s):  
Hugh G. Nimmo ◽  
Janet Laird

The core of the plant circadian clock involves multiple interlocking gene expression loops and post-translational controls along with inputs from light and metabolism. The complexity of the interactions is such that few specific functions can be ascribed to single components. In previous work, we reported differences in the operation of the clocks in Arabidopsis shoots and roots, including the effects of mutations of key clock components. Here, we have used luciferase imaging to study prr7 mutants expressing CCA1::LUC and GI::LUC markers. In mature shoots expressing CCA1::LUC, loss of PRR7 radically altered behaviour in light:dark cycles and caused loss of rhythmicity in constant light but had little effect on roots. In contrast, in mature plants expressing GI::LUC, loss of PRR7 had little effect in light:dark cycles but in constant light increased the circadian period in shoots and reduced it in roots. We conclude that most or all of the circadian input to the CCA1 promoter in shoots is mediated by PRR7 and that loss of PRR7 has organ-specific effects. The results emphasise the differences in operation of the shoot and root clocks, and the importance of studying clock mutants in both light:dark cycles and constant light.


Author(s):  
Stéphane Schmitt

The problem of the repeated parts of organisms was at the center of the biological sciences as early as the first decades of the 19th century. Some concepts and theories (e.g., serial homology, unity of plan, or colonial theory) introduced in order to explain the similarity as well as the differences between the repeated structures of an organism were reused throughout the 19th and the 20th century, in spite of the fundamental changes during this long period that saw the diffusion of the evolutionary theory, the rise of experimental approaches, and the emergence of new fields and disciplines. Interestingly, this conceptual heritage was at the core of any attempt to unify the problems of inheritance, development, and evolution, in particular in the last decades, with the rise of “evo-devo.” This chapter examines the conditions of this theoretical continuity and the challenges it brings out for the current evolutionary sciences.


Genetics ◽  
1976 ◽  
Vol 82 (1) ◽  
pp. 9-17 ◽  
Author(s):  
Jerry F Feldman ◽  
Marian N Hoyle

ABSTRACT A fourth mutant of Neurospora crassa, designated frq-4, has been isolated in which the period length of the circadian conidiation rhythm is shortened to 19.3 ± 0.3 hours. This mutant is tightly linked to the three previously isolated frq mutants, and all four map to the right arm of linkage group VII about 10 map units from the centromere. Complementation tests suggest, but do not prove, that all four mutations are allelic, since each of the four mutants is co-dominant with the frq  + allele—i.e., heterokaryons have period lengths intermediate between the mutant and wild-type—and since heterokaryons between pairs of mutants also have period lengths intermediate between those of the two mutants.


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