Direct Interaction between Ras Homolog Enriched in Brain and FK506 Binding Protein 38 in Cashmere Goat Fetal Fibroblast Cells

Wang, Xiaojing; Wang, Yanfeng; Xu, Zheng; Hao, Xiang; Liang, Yan; Wu, Manlin; Wang, Xiao; Wang, Zhigang

doi:10.5713/ajas.2014.14145

Cited by 6 publications

(8 citation statements)

References 33 publications

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“…In previous studies, we have confirmed the interactions between Rheb and FKBP38 experimentally in goat cells (Wang et al, 2014). In the present study, we showed that KBP38 gene-silencing induced phosphorylation of mTOR, S6, and 4EBP1.…”

Section: Discussionsupporting

confidence: 74%

Silencing FKBP38 gene by siRNA induces activation of mTOR signaling in goat fetal fibroblasts

Fu¹,

Zheng²,

He³

et al. 2015

Genet. Mol. Res.

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Section: Discussionsupporting

confidence: 74%

Silencing FKBP38 gene by siRNA induces activation of mTOR signaling in goat fetal fibroblasts

Fu¹,

Zheng²,

He³

et al. 2015

Genet. Mol. Res.

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“…Then, equivalent amounts of protein lysate were incubated with anti-4EBP1 (1:50) or anti-His (1:50) (negative control) and inactive resin (negative control) in Millipore catch-and-release spin columns. Equivalent amounts of protein lysate were also evaluated by Western blot as a positive control for 4EBP1 and PPARγ, per reported methods. , …”

Section: Methodsmentioning

confidence: 99%

The mTORC1/4EBP1/PPARγ Axis Mediates Insulin-Induced Lipogenesis by Regulating Lipogenic Gene Expression in Bovine Mammary Epithelial Cells

Guo

Cheng

Feng

et al. 2019

J. Agric. Food Chem.

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4EBP1 is a chief downstream factor of mTORC1, and PPARγ is a key lipogenesis-related transcription factor. mTORC1 and PPARγ are associated with lipid metabolism. However, it is unknown which effector protein connects mTORC1 and PPARγ. This study investigated the interaction between 4EBP1 with PPARγ as part of the underlying mechanism by which insulin-induced lipid synthesis and secretion are regulated by mTORC1 in primary bovine mammary epithelial cells (pBMECs). Rapamycin, a specific inhibitor of mTORC1, downregulated 4EBP1 phosphorylation and the expression of PPARγ and the following lipogenic genes: lipin 1, DGAT1, ACC, and FAS. Rapamycin also decreased the levels of intracellular triacylglycerol (TAG); 10 types of fatty acid; and the accumulation of TAG, palmitic acid (PA), and stearic acid (SA) in the cell culture medium. Inactivation of mTORC1 by shRaptor or shRheb attenuated the synthesis and secretion of TAG and PA. In contrast, activation of mTORC1 by Rheb overexpression promoted 4EBP1 phosphorylation and PPARγ expression and upregulated the mRNA and protein levels of lipin 1, DGAT1, ACC, and FAS, whereas the levels of intracellular and extracellular TAG, PA, and SA also rose. Further, 4EBP1 interacted directly with PPARγ. Inactivation of mTORC1 by shRaptor prevented the nuclear location of PPARγ. These results demonstrate that mTORC1 regulates lipid synthesis and secretion by inducing the expression of lipin 1, DGAT1, ACC, and FAS, which is likely mediated by the 4EBP1/PPARγ axis. This finding constitutes a novel mechanism by which lipid synthesis and secretion are regulated in pBMECs.

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“…An 1147 bp fragment of SQSTM1/p62 gene cDNA was subcloned into pGADT7 Vector (Clontech Laboratorie, Inc.) to generate the pGADT7-SQSTM1/p62 plasmid (expressing the prey protein GAL4AD-SQSTM1/p62). The pGBKT7-FKBP38 (expressing the bait protein GAL4BD-FKBP38) was described previously (Wang et al 2014). The plasmids pGADT7-SQSTM1/p62 and pGBKT7-FKBP38 were co-transferred into the Y2HGold yeast strain.…”

Section: Yeast Two-hybrid Analysis Of Protein Interactionmentioning

confidence: 99%

SQSTM1/p62 interacts with FKBP38 and regulates cell cycle in Cashmere goat foetal fibroblasts

Wang

Zhao

et al. 2018

Journal of Applied Animal Research

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SQSTM1 (sequestosome 1, also known as p62) is a multifunctional scaffold protein implicated in diverse cell physiology processes, such as autophagy, cell signalling, and protein turnover. FKBP38 (FK506-binding protein 38) is a member of FKBPs family and plays a key role in various cellular processes, including signalling transduction, embryonic development and apoptosis. In order to explore the role of SQSTM1/p62 gene in cell-cycle progression and proliferation of goat foetal fibroblast (GFbs), SQSTM1/ p62 gene was cloned and characterized. Furthermore, the yeast two-hybrid screening system was used to identify the interaction between SQSTM1/p62 and FKBP38. The results suggested that SQSTM1/p62 directly interacts with FKBP38. Overexpression vector pIRES-EGFP-SQSTM1/p62 and shRNA eukaryotic expression vector pRNAT-U6.1-shSQSTM1/p62 which harboured siRNA targeting the SQSTM1/p62 mRNA were constructed. The overexpression of SQSTM1/p62 gene in GFbs significantly increase the Sphase cells compared with control cells (p < .05). Furthermore, SQSTM1/p62 gene silencing in GFbs leads to a significant decrease of S-phase cells and cell cycle arrest compared with control cells (p < .05). These data indicate that SQSTM1/p62 gene plays an important role in cell-cycle and proliferation of Cashmere GFbs.

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Direct Interaction between Ras Homolog Enriched in Brain and FK506 Binding Protein 38 in Cashmere Goat Fetal Fibroblast Cells

Cited by 6 publications

References 33 publications

Silencing FKBP38 gene by siRNA induces activation of mTOR signaling in goat fetal fibroblasts

Silencing FKBP38 gene by siRNA induces activation of mTOR signaling in goat fetal fibroblasts

The mTORC1/4EBP1/PPARγ Axis Mediates Insulin-Induced Lipogenesis by Regulating Lipogenic Gene Expression in Bovine Mammary Epithelial Cells

SQSTM1/p62 interacts with FKBP38 and regulates cell cycle in Cashmere goat foetal fibroblasts

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