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dc.contributor.authorChen, Liangen_US
dc.date.accessioned2016-09-24T08:00:38Z
dc.date.available2016-09-24T08:00:38Z
dc.date.issued2016-09-23en_US
dc.identifier.othervt_gsexam:8853en_US
dc.identifier.urihttp://hdl.handle.net/10919/73037
dc.description.abstractThe key transcription factor sterol regulatory element binding protein-1 (SREBP1) plays a central role in milk fat synthesis. SREBP1 stimulates the transcription of genes encoding lipogenic enzymes. The overall objective of these studies was to investigate the mechanisms of SREBP1 regulation by nutrients. In the first study, chromatin immunoprecipitation (ChIP) accompanied with deep-sequencing was employed to investigate the potential sterol regulatory elements (SRE) in the promoter of SREBP1-target genes. The SRE in three known SREBP1-target genes SREBP1, fatty acid synthase (FASN) and stearoyl-CoA desaturase (SCD) were first validated in a bovine mammary epithelial cell line (MacT) and in bovine mammary tissues. At least one or two SRE binding sites in 24 selected lipogenic genes were identified within 50,000 base pair to the 5'-transcription start site through ChIP-seq. The genes closest to the highest enriched peaks were involved in cell integrity, defense or signal transduction whereas lipogenic genes were not among the top enrichment leading to the questions about the success of the ChIP. The second study was conducted to determine the effect of t10, c12-conjugated linoleic acid (CLA) on insulin induced gene-1 (Insig1), an endoplasmic reticulum (ER) protein that anchors SREBP1 and prevents proteolytic activation of SREBP1. MacT cells were treated with increasing levels of t10, c12-CLA. High concentration of t10, c12-CLA inhibited Insig1 degradation therefore decreased SREBP1 maturation. Furthermore, immunoprecipitation (IP) confirmed that t10, c12-CLA reduced Insig1 proteasomal iv degradation by disrupting the interaction between Insig1 and UBX domain-containing protein 8 (Ubxd8), which is part of a degradation complex that removes Insig1 from the ER. In the third study, three potential regulators of SREBP1 activation and their pathways were investigated in insulin, t10, c12-CLA or glucose treated MacT cells. Insulin-induced mammalian target of rapamycin (mTOR) signaling stimulated lipogenesis via activation of SREBP1 and the stimulatory effect was based on the regulation on cAMP response element binding protein coactivator 2 (CRTC2) phosphorylation, Lipin1 translocation and glycogen synthase kinase-3 (GSK3)-dependent proteasomal degradation. t10, c12-CLA inhibited SREBP1 through AMP-activated protein kinase (AMPK) phosphorylation, a key protein kinase in energy homeostasis. Glucose stabilized the SREBP1 chaperone protein SCAP and facilitated SREBP1 activation. Overall, SREBP1 activation is under specific regulation of t10, c12-CLA and interacts with multiple major cellular signaling pathways in response to hormonal stimulation and nutrient availability.en_US
dc.format.mediumETDen_US
dc.publisherVirginia Techen_US
dc.rightsThis Item is protected by copyright and/or related rights. Some uses of this Item may be deemed fair and permitted by law even without permission from the rights holder(s), or the rights holder(s) may have licensed the work for use under certain conditions. For other uses you need to obtain permission from the rights holder(s).en_US
dc.subjectSREBP1en_US
dc.subjectt10en_US
dc.subjectc12-CLAen_US
dc.subjectmTORen_US
dc.subjectInsig1en_US
dc.titleRegulation of sterol regulatory element binding protein-1 in bovine mammary epithelial cellsen_US
dc.typeDissertationen_US
dc.contributor.departmentDairy Scienceen_US
dc.description.degreePHDen_US
thesis.degree.namePHDen_US
thesis.degree.leveldoctoralen_US
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen_US
thesis.degree.disciplineAnimal Sciences, Dairyen_US
dc.contributor.committeechairCorl, Benjamin Aen_US
dc.contributor.committeememberFerreira, Gonzaloen_US
dc.contributor.committeememberJiang, Honglinen_US
dc.contributor.committeememberAkers, Robert Men_US


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