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Nuclear envelope protein MAN1 regulates clock through BMAL1.

Lin ST, Zhang L, Lin X, Zhang LC, Garcia VE, Tsai CW, Ptáček L, Fu YH - Elife (2014)

Bottom Line: Our knowledge of these components and pathways is far from exhaustive.In recent decades, the nuclear envelope has emerged as a global gene regulatory machine, although its role in circadian regulation has not been explored.Our results establish a novel connection between the nuclear periphery and circadian rhythmicity, therefore bridging two global regulatory systems that modulate all aspects of bodily functions.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University of California, San Francisco, San Francisco, United States.

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Domain-specific interactions between MAN1 and the hBMAL1 promoter.(A) Schematic representation of the indicated constructs generated from the 3.4 kb hBMAL1 promoter and cloned into the luciferase reporter vector pGL3. Histogram of luciferase activity in HEK293 cells transfected with the deleted hBMAL1 promoter constructs in the absence or presence of MAN1 expression vectors. Cells transfected for 48 hr and relative luciferase activities measured in extracts and normalized to Renilla luciferase activities. Activities (relative luciferase activity) are shown on the y-axis. Values are means ± SEM, n = 3, Student's t test, **p < 0.01, ***p < 0.001 compared to control. (B) Schematic representation of the deletion constructs generated from the MAN1 expressing construct. (C) Sequences of constructs with point mutations generated from the MAN1 expressing construct.DOI:http://dx.doi.org/10.7554/eLife.02981.019
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fig8s1: Domain-specific interactions between MAN1 and the hBMAL1 promoter.(A) Schematic representation of the indicated constructs generated from the 3.4 kb hBMAL1 promoter and cloned into the luciferase reporter vector pGL3. Histogram of luciferase activity in HEK293 cells transfected with the deleted hBMAL1 promoter constructs in the absence or presence of MAN1 expression vectors. Cells transfected for 48 hr and relative luciferase activities measured in extracts and normalized to Renilla luciferase activities. Activities (relative luciferase activity) are shown on the y-axis. Values are means ± SEM, n = 3, Student's t test, **p < 0.01, ***p < 0.001 compared to control. (B) Schematic representation of the deletion constructs generated from the MAN1 expressing construct. (C) Sequences of constructs with point mutations generated from the MAN1 expressing construct.DOI:http://dx.doi.org/10.7554/eLife.02981.019

Mentions: Since MAN1 does not execute its function through RORE, we investigated the promoter region of BMAL1 to determine what is necessary for the enhancing effect of MAN1. A series of deletion constructs of the BMAL1 promoter were generated for luciferase assays and a 900 bp region (−795 ∼ +106) was identified to be the region harboring the necessary DNA sequence for the regulatory effect of MAN1 on BMAL1 (Figure 8A, Figure 8—figure supplement 1A).10.7554/eLife.02981.018Figure 8.MAN1 binds to the BMAL1 promoter to enhance its transcription.


Nuclear envelope protein MAN1 regulates clock through BMAL1.

Lin ST, Zhang L, Lin X, Zhang LC, Garcia VE, Tsai CW, Ptáček L, Fu YH - Elife (2014)

Domain-specific interactions between MAN1 and the hBMAL1 promoter.(A) Schematic representation of the indicated constructs generated from the 3.4 kb hBMAL1 promoter and cloned into the luciferase reporter vector pGL3. Histogram of luciferase activity in HEK293 cells transfected with the deleted hBMAL1 promoter constructs in the absence or presence of MAN1 expression vectors. Cells transfected for 48 hr and relative luciferase activities measured in extracts and normalized to Renilla luciferase activities. Activities (relative luciferase activity) are shown on the y-axis. Values are means ± SEM, n = 3, Student's t test, **p < 0.01, ***p < 0.001 compared to control. (B) Schematic representation of the deletion constructs generated from the MAN1 expressing construct. (C) Sequences of constructs with point mutations generated from the MAN1 expressing construct.DOI:http://dx.doi.org/10.7554/eLife.02981.019
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150126&req=5

fig8s1: Domain-specific interactions between MAN1 and the hBMAL1 promoter.(A) Schematic representation of the indicated constructs generated from the 3.4 kb hBMAL1 promoter and cloned into the luciferase reporter vector pGL3. Histogram of luciferase activity in HEK293 cells transfected with the deleted hBMAL1 promoter constructs in the absence or presence of MAN1 expression vectors. Cells transfected for 48 hr and relative luciferase activities measured in extracts and normalized to Renilla luciferase activities. Activities (relative luciferase activity) are shown on the y-axis. Values are means ± SEM, n = 3, Student's t test, **p < 0.01, ***p < 0.001 compared to control. (B) Schematic representation of the deletion constructs generated from the MAN1 expressing construct. (C) Sequences of constructs with point mutations generated from the MAN1 expressing construct.DOI:http://dx.doi.org/10.7554/eLife.02981.019
Mentions: Since MAN1 does not execute its function through RORE, we investigated the promoter region of BMAL1 to determine what is necessary for the enhancing effect of MAN1. A series of deletion constructs of the BMAL1 promoter were generated for luciferase assays and a 900 bp region (−795 ∼ +106) was identified to be the region harboring the necessary DNA sequence for the regulatory effect of MAN1 on BMAL1 (Figure 8A, Figure 8—figure supplement 1A).10.7554/eLife.02981.018Figure 8.MAN1 binds to the BMAL1 promoter to enhance its transcription.

Bottom Line: Our knowledge of these components and pathways is far from exhaustive.In recent decades, the nuclear envelope has emerged as a global gene regulatory machine, although its role in circadian regulation has not been explored.Our results establish a novel connection between the nuclear periphery and circadian rhythmicity, therefore bridging two global regulatory systems that modulate all aspects of bodily functions.

View Article: PubMed Central - PubMed

Affiliation: Department of Neurology, University of California, San Francisco, San Francisco, United States.

Show MeSH
Related in: MedlinePlus