• Title/Summary/Keyword: clock mutant

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Proteomic Analysis of Circadian Clock Mutant Mice

  • Lee Joon-Woo;Kim Han-Gyu;Bae Kiho
    • Biomedical Science Letters
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    • v.11 no.4
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    • pp.493-501
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    • 2005
  • Circadian rhythms, time on a scale of about 24 hours, are present in a number of organisms including animals, plants, and bacteria. The control of the biochemical, physiological and behavioral processes is regulated by endogenous clocks in the suprachiasmatic nucleus (SCN). At the core of this timing mechanism is molecular machinery that are present both in the brain and in the peripheral tissues throughout the body, and even in a single cultured cell. In this study, we performed two-dimensional gel electrophoresis to figure out any correlation between protein expression patterns and the requirement of two canonical clock proteins, either mPER1 or mPER2, by comparing global protein expression profiles in livers from wildtype or mPer1/mPer2 double mutant mice. We could identify several differentially expressed protein candidates with respect to time and genotypes. Further analysis of these candidate proteins in detail in vivo will lead us to the better understanding of how circadian clock functions in mammals.

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Possible involvement of temperature-entrainable timing system in arrhythmic mutant flies in Drosophila melanogaster

  • Yoshii, Taishi;Tomioka, Kenji
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.240-242
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    • 2002
  • In Drosophila melanogaster, it is known that the circadian clock consists of an autoregulatory feedback loop, which includes so-called clock genes, such as per, tim, dClk and cyc and produces periodical expression of per. It is recently suggested, however, that the circadian oscillation without the rhythmical expression of per is involved in the regulation of circadian locomotor rhythms. In the present study, we examined the existence and the property of the possible per-less oscillation using arrhythmic clock mutant flies carrying per$^{01}$, tim$^{01}$, dClk$^{Jrk}$ or cyc$^{01}$. When temperature cycles consisting of 25$^{\circ}$C and 30$^{\circ}$C with varying periods (T = 8~32 hr) were given, they showed rhythms synchronizing with the given cycle under constant darkness (DD). per$^{01}$ and tim$^{01}$ flies always showed a peak around 7 hr after the onset of thermophase irrespective of Ts of temperature cycles, while dClk$^{Jrk}$ and cyc$^{01}$ flies did not. In addition, several days were necessary to establish a clear temperature entrainment in per$^{01}$ and tim$^{01}$ flies, when they were transferred from a constant temperature to a temperature cycle under DD. These results suggest that per$^{01}$ and tim$^{01}$ flies have a temperature-entrainable weak oscillatory mechanism. The fact that dClk$^{Jrk}$ and cyc$^{01}$ flies did not show any sign of the endogenous oscillation suggests that the per-less oscillatory mechanism may require CLK and CYC.

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Differential Effects of Two Period Genes on the Physiology and Proteomic Profiles of Mouse Anterior Tibialis Muscles

  • Bae, Kiho;Lee, Kisoo;Seo, Younguk;Lee, Haesang;Kim, Dongyong;Choi, Inho
    • Molecules and Cells
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    • v.22 no.3
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    • pp.275-284
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    • 2006
  • The molecular components that generate and maintain circadian rhythms of physiology and behavior in mammals are present both in the brain (suprachiasmatic nucleus; SCN) and in peripheral tissues. Examination of mice with targeted disruptions of either mPer1 or mPer2 has shown that these two genes have key roles in the SCN circadian clock. Here we show that loss of the clock gene mPer2 affects forced locomotor performance in mice without altering muscle contractility. A proteomic analysis revealed that the anterior tibialis muscles of the mPer2 knockout mice had higher levels of glycolytic enzymes such as triose phosphate isomerase and enolase than those of either the wild type or mPer1 knockout mice. In addition, the level of expression of HSP90 in the mPer2 mutant mice was also significantly higher than in wildtype mice. These results suggest that the reduced locomotor endurance of the mPer2 knockout mice reflects a greater dependence on anaerobic metabolism under stress conditions, and that the two canonical clock genes, mPer1 and mPer2, play distinct roles in the physiology of skeletal muscle.

Diversification of the molecular clockwork for tissue-specific function: insight from a novel Drosophila Clock mutant homologous to a mouse Clock allele

  • Cho, Eunjoo;Lee, Euna;Kim, Eun Young
    • BMB Reports
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    • v.49 no.11
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    • pp.587-589
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    • 2016
  • The circadian clock system enables organisms to anticipate the rhythmic environmental changes and to manifest behavior and physiology at advantageous times of the day. Transcriptional/translational feedback loop (TTFL) is the basic feature of the eukaryotic circadian clock and is based on the rhythmic association of circadian transcriptional activator and repressor. In Drosophila, repression of dCLOCK/CYCLE (dCLK/CYC) mediated transcription by PERIOD (PER) is critical for inducing circadian rhythms of gene expression. Pacemaker neurons in the brain control specific circadian behaviors upon environmental timing cues such as light and temperature cycle. We show that amino acids 657-707 of dCLK are important for the transcriptional activation and the association with PER both in vitro and in vivo. Flies expressing dCLK lacking AA657-707 in $Clk^{out}$ genetic background, homologous to the mouse Clock allele where exon 19 region is deleted, display pacemaker-neuron-dependent perturbation of the molecular clockwork. The molecular rhythms in light-cycle-sensitive pacemaker neurons such as ventral lateral neurons ($LN_vs$) were significantly disrupted, but those in temperature-cycle-sensitive pacemaker neurons such as dorsal neurons (DNs) were robust. Our results suggest that the dCLK-controlled TTFL diversify in a pacemaker-neuron-dependent manner which may contribute to specific functions such as different sensitivities to entraining cues.

Antibiotic Biosynthesis in bldA-like Mutant of Strptomyces coelicolor (Streptomyces coelicolor blAA-like Mutant에서의 항생물질 생합성)

  • Park, Unn-Mee
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.70-77
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    • 1994
  • The author isolated 7 mutant candidates which mapped around cysA (which was 10 o'clock). They were divided into two groups. One of them was located counterclockwise to cysA, and the other was clockwise to cysA. Since bldA was mapped counterclockwise to cysA, the candidate which mapped counterclockwise to cysA was transduced with phage containing wild type bldA gene clone. The candidates might be the alleles of bldA, because they were complemented by bldA clone. However some of such mutants sporulated very well and developed as much pigment as wild type on rich media plate. Their phenotype was not like bld mutant at all on such conditions. There were real antibiotics gene expressions, since transcriptional reporter gene xylE had shown high activities. Majority of the bldA like mutants showed act gene expressions when they were transformed with high copy number plasmid containing actII-ORF4.

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Photoperiodic modulation of insect circadian rhythms

  • Tomioka, Kenji;Uwozumi, Kouzo;Koga, Mika
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.9-12
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    • 2002
  • Circadian rhythms can be seen in a variety of physiological functions in insects. Light is a powerful zeitgeber not only synchronizing but also modulating the rhythm to adjust insect's temporal structure to seasonal changes in the environmental cycle. There are two general effects of the length of light phase within 24 hr light cycles on the circadian rhythms, i.e., the modulation of free-running period and the waveform. Since the photoperiodic modulation of the free-running period is induced even in the clock mutant flies, per$\^$s/, the free-running period is not fully determined genetically. In crickets, the ratio of activity (a) and rest phase (p) under the constant darkness (DD) is clearly dependent on the photoperiod under which they have been kept. When experienced the longer photoperiod it becomes smaller. The magnitude of change in a/p-ratio is dependent on the number of cycles they experienced. The neuronal activity of the optic lobe in DD shows the a/p-ratio changing with the preceding photoperiod. These data suggest that a single circadian pacemaker stores and maintains the photoperiodic information and that there is a system that accumulates the effects of single photoperiod to cause greater effects.

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Genetic Analysis of absR, a new abs locus of Streptomyces coelicolor

  • Park, Uhn-Mee;Suh, Joo-Won;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.10 no.2
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    • pp.169-175
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    • 2000
  • The filamentous soil bacterium Streptomyces coelicolor is known to produce four distinct antibiotics. The simultaneous global regulation for the biosynthesis of those four antibiotics was previously confirmed by absA and absB mutations that blocked all four antibiotics' biosynthesis without influencing their morphological differentiation. To study the complex regulatory cascade that controls the secondary metabolism in Streptomyces, a new abs-like mutation was characterized. namely absR, which is slightly leaky on a complete R2YE medium, yet tight on a minimal medium. A genetic analysis of the absR locus indicated that it is located at 10 o'clock on the genetic map, near the site of absA. A cloned copy of the absA gene that encoded bacterial two-component regulatory kinases did not restore antibiotic biosyntheis to the absR mutant. Accordingly, it is proposed that absR is another abs-type mutation which is less tight than the previously identified absA or absB mutations income medium conditions, and can be used to characterize another global regulatory gene for secondary metabolete formation in S. coelicolor.

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Circadin Gibberellins production in Sorghum (수수에서 식물호르몬 지베렐린의 circadian 리듬)

  • 이인중;김길웅;김경민
    • Journal of Life Science
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    • v.7 no.3
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    • pp.205-211
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    • 1997
  • The possibility of circadian production of plant hormone gibberellin (GA0 was examined in phytochrome B mutant (plyB-1) and wild-type sorghum. GA$_{12}$, GA$_{20}$ and GA$_{1}$ levels were found to cycle circadianly in both phyB-1 and wild-type. The periods (33 h) of GA$_{20}$ and GA$_{1}$ cycling in constant light were longer than normal photoperiods in both genotypes and typical average free running periods in plants of 22 to 28 h. The biological clock was thus shown to function properly in phyB-1. However, circadian regulation of GAs productions were not clear as compared to circadian ethylene regulation reported by Lee (1996). Although, in sorghum, EOD FR treatment hasten floral inititation, the differences in GA concentrations between treatments and untreated control were generally less dramatic than expected. Thus, it can be concluded that FR does not act primarily by changing absolute levels of GAs but rather by increasing flowering responsiveness to GAs.

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