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Transcriptional interference by antisense RNA is required for circadian clock function.

Xue Z, Ye Q, Anson SR, Yang J, Xiao G, Kowbel D, Glass NL, Crosthwaite SK, Liu Y - Nature (2014)

Bottom Line: Natural antisense RNAs are found in a wide range of eukaryotic organisms.Moreover, our results suggest that antisense transcription inhibits sense expression by mediating chromatin modifications and premature termination of transcription.Taken together, our results establish antisense transcription as an essential feature in a circadian system and shed light on the importance and mechanism of antisense action.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390, USA.

ABSTRACT
Eukaryotic circadian oscillators consist of negative feedback loops that generate endogenous rhythmicities. Natural antisense RNAs are found in a wide range of eukaryotic organisms. Nevertheless, the physiological importance and mode of action of most antisense RNAs are not clear. frequency (frq) encodes a component of the Neurospora core circadian negative feedback loop, which was thought to generate sustained rhythmicity. Transcription of qrf, the long non-coding frq antisense RNA, is induced by light, and its level oscillates in antiphase to frq sense RNA. Here we show that qrf transcription is regulated by both light-dependent and light-independent mechanisms. Light-dependent qrf transcription represses frq expression and regulates clock resetting. Light-independent qrf expression, on the other hand, is required for circadian rhythmicity. frq transcription also inhibits qrf expression and drives the antiphasic rhythm of qrf transcripts. The mutual inhibition of frq and qrf transcription thus forms a double negative feedback loop that is interlocked with the core feedback loop. Genetic and mathematical modelling analyses indicate that such an arrangement is required for robust and sustained circadian rhythmicity. Moreover, our results suggest that antisense transcription inhibits sense expression by mediating chromatin modifications and premature termination of transcription. Taken together, our results establish antisense transcription as an essential feature in a circadian system and shed light on the importance and mechanism of antisense action.

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qrf expression is required for circadian rhythmicities. (a) Northern blot analysis showing the levels of frq and qrf with/without 1×10-3 M QA (0.1% glucose) in LL. Right panel shows the densitometric analyses of the results. Error bars are SD. (b) Race tube analyses of the indicated strains (0.1% glucose with different concentrations of QA) in DD. (c) Luciferase reporter assay showing the normalized frq promoter activity after two days in DD.
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Figure 12: qrf expression is required for circadian rhythmicities. (a) Northern blot analysis showing the levels of frq and qrf with/without 1×10-3 M QA (0.1% glucose) in LL. Right panel shows the densitometric analyses of the results. Error bars are SD. (b) Race tube analyses of the indicated strains (0.1% glucose with different concentrations of QA) in DD. (c) Luciferase reporter assay showing the normalized frq promoter activity after two days in DD.

Mentions: Similar levels of qrf transcripts seen in the frqKI(WT) and frqKI(qLRE mut) strains at DD24 (Extended Data Figure 2a) indicate that qLRE does not regulate qrf expression in DD. In a strain (frq10;frq.aq) in which the promoter of qrf was replaced with the quinic acid (QA)-inducible qa-2 promoter, qrf expression was abolished in the absence of QA (Figure 2a). On addition of QA, qrf was induced but frq levels were significantly reduced, further indicating repression of frq by qrf.


Transcriptional interference by antisense RNA is required for circadian clock function.

Xue Z, Ye Q, Anson SR, Yang J, Xiao G, Kowbel D, Glass NL, Crosthwaite SK, Liu Y - Nature (2014)

qrf expression is required for circadian rhythmicities. (a) Northern blot analysis showing the levels of frq and qrf with/without 1×10-3 M QA (0.1% glucose) in LL. Right panel shows the densitometric analyses of the results. Error bars are SD. (b) Race tube analyses of the indicated strains (0.1% glucose with different concentrations of QA) in DD. (c) Luciferase reporter assay showing the normalized frq promoter activity after two days in DD.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4214883&req=5

Figure 12: qrf expression is required for circadian rhythmicities. (a) Northern blot analysis showing the levels of frq and qrf with/without 1×10-3 M QA (0.1% glucose) in LL. Right panel shows the densitometric analyses of the results. Error bars are SD. (b) Race tube analyses of the indicated strains (0.1% glucose with different concentrations of QA) in DD. (c) Luciferase reporter assay showing the normalized frq promoter activity after two days in DD.
Mentions: Similar levels of qrf transcripts seen in the frqKI(WT) and frqKI(qLRE mut) strains at DD24 (Extended Data Figure 2a) indicate that qLRE does not regulate qrf expression in DD. In a strain (frq10;frq.aq) in which the promoter of qrf was replaced with the quinic acid (QA)-inducible qa-2 promoter, qrf expression was abolished in the absence of QA (Figure 2a). On addition of QA, qrf was induced but frq levels were significantly reduced, further indicating repression of frq by qrf.

Bottom Line: Natural antisense RNAs are found in a wide range of eukaryotic organisms.Moreover, our results suggest that antisense transcription inhibits sense expression by mediating chromatin modifications and premature termination of transcription.Taken together, our results establish antisense transcription as an essential feature in a circadian system and shed light on the importance and mechanism of antisense action.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, The University of Texas Southwestern Medical Center, 5323 Harry Hines Boulevard, Dallas, Texas 75390, USA.

ABSTRACT
Eukaryotic circadian oscillators consist of negative feedback loops that generate endogenous rhythmicities. Natural antisense RNAs are found in a wide range of eukaryotic organisms. Nevertheless, the physiological importance and mode of action of most antisense RNAs are not clear. frequency (frq) encodes a component of the Neurospora core circadian negative feedback loop, which was thought to generate sustained rhythmicity. Transcription of qrf, the long non-coding frq antisense RNA, is induced by light, and its level oscillates in antiphase to frq sense RNA. Here we show that qrf transcription is regulated by both light-dependent and light-independent mechanisms. Light-dependent qrf transcription represses frq expression and regulates clock resetting. Light-independent qrf expression, on the other hand, is required for circadian rhythmicity. frq transcription also inhibits qrf expression and drives the antiphasic rhythm of qrf transcripts. The mutual inhibition of frq and qrf transcription thus forms a double negative feedback loop that is interlocked with the core feedback loop. Genetic and mathematical modelling analyses indicate that such an arrangement is required for robust and sustained circadian rhythmicity. Moreover, our results suggest that antisense transcription inhibits sense expression by mediating chromatin modifications and premature termination of transcription. Taken together, our results establish antisense transcription as an essential feature in a circadian system and shed light on the importance and mechanism of antisense action.

Show MeSH