Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression
Objective: Leptin gene expression is highly correlated with cellular lipid content in adipocytes but the transcriptional mechanisms controlling leptin expression in vivo are poorly understood. In this report, we set out to identify cis- and trans-regulatory elements controlling leptin expression. Me...
Main Authors: | , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2015-05-01
|
Series: | Molecular Metabolism |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2212877815000277 |
id |
doaj-72ec5ea105124c3d955c7c35314feed2 |
---|---|
record_format |
Article |
spelling |
doaj-72ec5ea105124c3d955c7c35314feed22020-11-24T21:00:00ZengElsevierMolecular Metabolism2212-87782015-05-014539240510.1016/j.molmet.2015.02.002Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expressionYi-Hsueh Lu0Olof Stefan Dallner1Kivanc Birsoy2Gulya Fayzikhodjaeva3Jeffrey M. Friedman4Laboratory of Molecular Genetics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USALaboratory of Molecular Genetics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USALaboratory of Molecular Genetics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USALaboratory of Molecular Genetics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USALaboratory of Molecular Genetics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USAObjective: Leptin gene expression is highly correlated with cellular lipid content in adipocytes but the transcriptional mechanisms controlling leptin expression in vivo are poorly understood. In this report, we set out to identify cis- and trans-regulatory elements controlling leptin expression. Methods: Leptin-BAC luciferase transgenic mice combining with other computational and molecular techniques were used to identify transcription regulatory elements including a CCAAT-binding protein Nuclear Factor Y (NF-Y). The function of NF-Y in adipocyte was studied in vitro with 3T3-L1 cells and in vivo with adipocyte-specific knockout of NF-Y. Results: Using Leptin-BAC luciferase mice, we showed that DNA sequences between −22 kb and +8.8 kb can confer quantitative expression of a leptin reporter. Computational analysis of sequences and gel shift assays identified a 32 bp sequence (chr6: 28993820–2899385) consisting a CCAAT binding site for Nuclear Factor Y (NF-Y) and this was confirmed by a ChIP assay in vivo. A deletion of this 32 bp sequence in the −22 kb to +8.8 kb leptin-luciferase BAC reporter completely abrogates luciferase reporter activity in vivo. RNAi mediated knockdown of NF-Y interfered with adipogenesis in vitro and adipocyte-specific knockout of NF-Y in mice reduced expression of leptin and other fat specific genes in vivo. Further analyses of the fat specific NF-Y knockout revealed that these animals develop a moderately severe lipodystrophy that is remediable with leptin therapy. Conclusions: These studies advance our understanding of leptin gene expression and show that NF-Y controls the expression of leptin and other adipocyte genes and identifies a new form of lipodystrophy.http://www.sciencedirect.com/science/article/pii/S2212877815000277LeptinAdipocyteNF-YAdipogenesisTranscriptional regulation |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yi-Hsueh Lu Olof Stefan Dallner Kivanc Birsoy Gulya Fayzikhodjaeva Jeffrey M. Friedman |
spellingShingle |
Yi-Hsueh Lu Olof Stefan Dallner Kivanc Birsoy Gulya Fayzikhodjaeva Jeffrey M. Friedman Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression Molecular Metabolism Leptin Adipocyte NF-Y Adipogenesis Transcriptional regulation |
author_facet |
Yi-Hsueh Lu Olof Stefan Dallner Kivanc Birsoy Gulya Fayzikhodjaeva Jeffrey M. Friedman |
author_sort |
Yi-Hsueh Lu |
title |
Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression |
title_short |
Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression |
title_full |
Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression |
title_fullStr |
Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression |
title_full_unstemmed |
Nuclear Factor-Y is an adipogenic factor that regulates leptin gene expression |
title_sort |
nuclear factor-y is an adipogenic factor that regulates leptin gene expression |
publisher |
Elsevier |
series |
Molecular Metabolism |
issn |
2212-8778 |
publishDate |
2015-05-01 |
description |
Objective: Leptin gene expression is highly correlated with cellular lipid content in adipocytes but the transcriptional mechanisms controlling leptin expression in vivo are poorly understood. In this report, we set out to identify cis- and trans-regulatory elements controlling leptin expression.
Methods: Leptin-BAC luciferase transgenic mice combining with other computational and molecular techniques were used to identify transcription regulatory elements including a CCAAT-binding protein Nuclear Factor Y (NF-Y). The function of NF-Y in adipocyte was studied in vitro with 3T3-L1 cells and in vivo with adipocyte-specific knockout of NF-Y.
Results: Using Leptin-BAC luciferase mice, we showed that DNA sequences between −22 kb and +8.8 kb can confer quantitative expression of a leptin reporter. Computational analysis of sequences and gel shift assays identified a 32 bp sequence (chr6: 28993820–2899385) consisting a CCAAT binding site for Nuclear Factor Y (NF-Y) and this was confirmed by a ChIP assay in vivo. A deletion of this 32 bp sequence in the −22 kb to +8.8 kb leptin-luciferase BAC reporter completely abrogates luciferase reporter activity in vivo. RNAi mediated knockdown of NF-Y interfered with adipogenesis in vitro and adipocyte-specific knockout of NF-Y in mice reduced expression of leptin and other fat specific genes in vivo. Further analyses of the fat specific NF-Y knockout revealed that these animals develop a moderately severe lipodystrophy that is remediable with leptin therapy.
Conclusions: These studies advance our understanding of leptin gene expression and show that NF-Y controls the expression of leptin and other adipocyte genes and identifies a new form of lipodystrophy. |
topic |
Leptin Adipocyte NF-Y Adipogenesis Transcriptional regulation |
url |
http://www.sciencedirect.com/science/article/pii/S2212877815000277 |
work_keys_str_mv |
AT yihsuehlu nuclearfactoryisanadipogenicfactorthatregulatesleptingeneexpression AT olofstefandallner nuclearfactoryisanadipogenicfactorthatregulatesleptingeneexpression AT kivancbirsoy nuclearfactoryisanadipogenicfactorthatregulatesleptingeneexpression AT gulyafayzikhodjaeva nuclearfactoryisanadipogenicfactorthatregulatesleptingeneexpression AT jeffreymfriedman nuclearfactoryisanadipogenicfactorthatregulatesleptingeneexpression |
_version_ |
1716780699667660800 |