Tudor-SN Regulates Milk Synthesis and Proliferation of Bovine Mammary Epithelial Cells

Ao, Jinxia; Wei, Changyong; Yu, Shaohua; Lv, Wei; Lin, Ye; Cui, Yingjun; Gao, Xuejun

doi:10.3390/ijms161226212

Cited by 28 publications

(28 citation statements)

References 47 publications

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“…In addition, we found that miR-139 inhibited not only the activity of downstream signaling molecules of the GHR and IGF1R pathways, but also the total protein levels of key molecules of these pathways. This phenomenon has also been found by others [ 24 , 35 ]. These observations suggest that miR-139 likely inhibits synthesis of β-casein through GHR and IGF1R signaling in BMECs.…”

Section: Discussionsupporting

confidence: 88%

MiR-139 suppresses β-casein synthesis and proliferation in bovine mammary epithelial cells by targeting the GHR and IGF1R signaling pathways

et al. 2017

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BackgroundMicroRNAs have important roles in many biological processes. However, the role of miR-139 in healthy mammary gland remains unclear. The objective of this study was to investigate the effects of miR-139 on lactation in dairy cows.ResultsHere, we found that miR-139 was down-regulated in mid-lactation dairy cow mammary tissues compared with mid-pregnancy tissues. Then, we prioritized two of potential target genes of miR-139 in cow, growth hormone receptor (GHR) and type I insulin-like growth factor receptor (IGF1R) for further functional studies based on their roles in lactation processes. Dual luciferase reporter assays validated direct binding of miR-139 to the 3′- untranslated region (UTR) of GHR and IGF1R. Moreover, over-expression or silencing of miR-139 affected mRNA levels of GHR and IGF1R in cultured bovine mammary epithelial cells (BMECs). Furthermore, over-expression of miR-139 decreased protein levels of β-casein, proliferation in mammary epithelial cell, and the protein levels of IGF1R and key members of the GHR or IGF1R pathways as well, whereas silencing miR-139 produced the opposite result. Among these signal molecules, signal transducer and activator of transcription-5 (STAT5), protein kinase B (also known as AKT1), mammalian target of rapamycin (mTOR), and p70-S6 Kinase (p70S6K) are involed in β-casein synthesis, and Cyclin D1 is involved in cell proliferation. In addition, silencing GHR decreased protein levels of β-casein, IGF1R, and key members of the IGF1R pathway, whereas co-silencing miR-139 and GHR rescued the expression of GHR and reversed GHR silencing effects.ConclusionsOur results demonstrate that GHR and IGF1R are target genes of miR-139 in dairy cow. MiR-139 suppresses β-casein synthesis and proliferation in BMECs by targeting the GHR and IGF1R signaling pathways.

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Section: Discussionsupporting

confidence: 88%

MiR-139 suppresses β-casein synthesis and proliferation in bovine mammary epithelial cells by targeting the GHR and IGF1R signaling pathways

et al. 2017

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“…Primary BMECs were separated and cultured according to previously reported methods (Ao et al, ; L. Wang et al, ). BMECs were cultured in glass cell culture plates in an incubator at 37°C in the presence of 5% CO 2 .…”

Section: Methodsmentioning

confidence: 99%

“…), amino acids (Met, Leu, etc. ), glucose, and fatty acids promote milk fat synthesis through the SREBP-1c pathway, but the detailed mechanism is still unclear (Ao et al, 2015;Zhang et al, 2018).…”

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confidence: 99%

FABP5 is a critical regulator of methionine‐ and estrogen‐induced SREBP‐1c gene expression in bovine mammary epithelial cells

Zhou

et al. 2018

Journal Cellular Physiology

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The intracellular fatty acid-binding proteins (FABPs) are a well-conserved family that function as lipid chaperones. Ongoing studies are focused on identification of the mechanistic complexity and vast biological diversity of different isoforms of FABPs. However, the molecular mechanism of FABP5 in the regulation of milk fat synthesis in the mammary gland of dairy cows is still largely unknown. Here, we report that FABP5 acts as a critical regulator of terol response element-binding protein-1c (SREBP-1c) gene expression induced by methionine (Met) and estrogen (E2) in bovine mammary epithelial cells (BMECs). We observed that the expression of FABP5 was markedly higher in dairy cow mammary tissue during the lactating period than the puberty period and the dry period. FABP5 is located in the cytoplasm, and Met and E2 significantly increase the protein levels of FABP5 in BMECs. Using gene function study approaches, we revealed that FABP5 positively regulates SREBP-1c gene expression and promotes milk fat synthesis. We confirmed that FABP5 is required for Met- and E2-induced SREBP-1c gene expression and milk fat synthesis. We further uncovered that fatty acids are needed for FABP5-mediated SREBP-1c gene expression. Thus, our study demonstrates that FABP5 is a critical regulator of Met- and E2-induced SREBP-1c gene expression leading to milk fat synthesis.

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“…Then we performed western blot analysis to determine whether the phosphorylation of the Tudor‐SN affected the expression of the milk proteins and mTOR signaling pathway which has been confirmed to be a positive regulator to control milk protein synthesis and cell proliferation of BMECs (Wang et al, ; Zhang et al, ). Furthermore, Tudor‐SN has been verified to positively regulate milk synthesis through mTOR signaling pathway (Ao et al, ). Therefore, mTOR and β‐casein were used as the milk protein synthesis marker.…”

Section: Resultsmentioning

confidence: 99%

The phosphorylation of Tudor‐SN mediated by JNK is involved in the regulation of milk protein synthesis induced by prolactin in BMECs

Zhang

Wen

et al. 2018

Journal Cellular Physiology

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Tudor staphylococcal nuclease (Tudor-SN) is a multifunctional protein involved in a variety of cellular processes and plays a critical role in the regulation of gene expression. Recently, Tudor-SN was found to be upregulated in mammary epithelial cells during lactation in response to prolactin, which further to regulate milk protein synthesis. However, the detailed regulatory mechanism of Tudor-SN to milk protein still remains to be elucidated. In our study, we observed that the levels of Tudor-SN and phosphor-Tudor-SN (Thr103) were both enhanced upon prolactin stimulation. Immunofluorescence assays demonstrated that prolactin treatment facilitated the nuclear transport of Tudor-SN. Further study revealed that the phosphorylation of Tudor-SN was depended on activated JNK. Coimmunoprecipitation assays disclosed that Tudor-SN might be phosphorylated directly by JNK. Using gene mutation assays, we further discovered that mutation of Thr to Ala at site of 103 prevented the nuclear transport of Tudor-SN. Thus, these results suggested the essential mechanism of the activated Tudor-SN in milk protein regulation in response to prolactin, which may provide some new sights into improve milk protein production.

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Tudor-SN Regulates Milk Synthesis and Proliferation of Bovine Mammary Epithelial Cells

Cited by 28 publications

References 47 publications

MiR-139 suppresses β-casein synthesis and proliferation in bovine mammary epithelial cells by targeting the GHR and IGF1R signaling pathways

MiR-139 suppresses β-casein synthesis and proliferation in bovine mammary epithelial cells by targeting the GHR and IGF1R signaling pathways

FABP5 is a critical regulator of methionine‐ and estrogen‐induced SREBP‐1c gene expression in bovine mammary epithelial cells

The phosphorylation of Tudor‐SN mediated by JNK is involved in the regulation of milk protein synthesis induced by prolactin in BMECs

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