MAN1 promotes BMAL1 transcriptional activity.(A) Reduct

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. Show MeSH Major ARNTL Transcription Factors/genetics/metabolism Circadian Clocks/genetics Gene Expression Regulation* Membrane Proteins/genetics/metabolism Nuclear Proteins/genetics/metabolism Minor Animals Blotting, Western Cell Line, Tumor HEK293 Cells Humans Lamin Type B/genetics/metabolism Mice, Inbred C57BL Mice, Knockout Nuclear Envelope/metabolism Promoter Regions, Genetic/genetics Protein Binding RNA Interference Reverse Transcriptase Polymerase Chain Reaction Related in: MedlinePlus	© Copyright Policy - open-access Related In: Results - Collection License Show All Figures getmorefigures.php?uid=PMC4150126&req=5 fig6: MAN1 promotes BMAL1 transcriptional activity.(A) Reduction of MAN1 transcripts (13 nM siRNA) reduced Bmal1 promoter activity (n = 3, p < 0.001). (B) Over-expression of FLAG-tagged MAN1 (F-MAN1) enhanced Bmal1-Luc activity (n = 3, p < 0.001). (C) Over-expression of FLAG-tagged MAN1 (F-MAN1) enhanced luciferase activities driven by mBmal1 promoter (530 bp) or hBMAL1 promoter (3.4 kb) (n = 3, p < 0.05). Error bars represent SD.DOI:http://dx.doi.org/10.7554/eLife.02981.016 Mentions:* A luciferase reporter assay using HEK293 cells was used to further investigate the effect of MAN1 on BMAL1 transcription. MAN1 knockdown decreased Bmal1-Luc activity by 72% (Figure 6A), whereas overexpression of FLAG-MAN1 increased the luciferase activity by more than twofold vs cells transfected with empty vector (Figure 6B). Similar results were obtained with a longer, human BMAL1 promoter (Figure 6C). These data indicate that MAN1 may play a role in circadian regulation by activating the promoter of BMAL1.10.7554/eLife.02981.016Figure 6.MAN1 promotes BMAL1 transcriptional activity.

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.

Show MeSH

Related in: MedlinePlus

MAN1 promotes BMAL1 transcriptional activity.(A) Reduction of MAN1 transcripts (13 nM siRNA) reduced Bmal1 promoter activity (n = 3, *p < 0.001). (B) Over-expression of FLAG-tagged MAN1 (F-MAN1) enhanced Bmal1-Luc activity (n = 3, *p < 0.001). (C) Over-expression of FLAG-tagged MAN1 (F-MAN1) enhanced luciferase activities driven by mBmal1 promoter (530 bp) or hBMAL1 promoter (3.4 kb) (n = 3, *p < 0.05). Error bars represent SD.DOI:http://dx.doi.org/10.7554/eLife.02981.016

Related In: Results - Collection

License

Show All Figures

getmorefigures.php?uid=PMC4150126&req=5

fig6: MAN1 promotes BMAL1 transcriptional activity.(A) Reduction of MAN1 transcripts (13 nM siRNA) reduced Bmal1 promoter activity (n = 3, *p < 0.001). (B) Over-expression of FLAG-tagged MAN1 (F-MAN1) enhanced Bmal1-Luc activity (n = 3, *p < 0.001). (C) Over-expression of FLAG-tagged MAN1 (F-MAN1) enhanced luciferase activities driven by mBmal1 promoter (530 bp) or hBMAL1 promoter (3.4 kb) (n = 3, *p < 0.05). Error bars represent SD.DOI:http://dx.doi.org/10.7554/eLife.02981.016

Mentions: A luciferase reporter assay using HEK293 cells was used to further investigate the effect of MAN1 on BMAL1 transcription. MAN1 knockdown decreased Bmal1-Luc activity by 72% (Figure 6A), whereas overexpression of FLAG-MAN1 increased the luciferase activity by more than twofold vs cells transfected with empty vector (Figure 6B). Similar results were obtained with a longer, human BMAL1 promoter (Figure 6C). These data indicate that MAN1 may play a role in circadian regulation by activating the promoter of BMAL1.10.7554/eLife.02981.016Figure 6.MAN1 promotes BMAL1 transcriptional activity.