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Septin6 regulates cell growth and casein synthesis in dairy cow mammary epithelial cells via mTORC1 pathway

Published online by Cambridge University Press:  24 May 2019

Bin Li
Affiliation:
Institute of Animal Husbandry and Veterinary, Tibet Autonomous Regional Academy of Agricultural Sciences, Lhasa, Tibet, 850000, China State Key Laboratory of Hulless and Yak Germplasm Resources and Genetic Improvement, Lhasa, Tibet, 850000, China
Zhuzha Basang
Affiliation:
Institute of Animal Husbandry and Veterinary, Tibet Autonomous Regional Academy of Agricultural Sciences, Lhasa, Tibet, 850000, China
Lijun Hu
Affiliation:
Colleges of Life Science and Technology, Dalian University, Dalian Economic Technological Development Zone, Liaoning, 116622, China
Liu Liu
Affiliation:
Colleges of Life Science and Technology, Dalian University, Dalian Economic Technological Development Zone, Liaoning, 116622, China
Nan Jiang*
Affiliation:
Institute of Animal Husbandry and Veterinary, Tibet Autonomous Regional Academy of Agricultural Sciences, Lhasa, Tibet, 850000, China Colleges of Life Science and Technology, Dalian University, Dalian Economic Technological Development Zone, Liaoning, 116622, China
*
Author for correspondence: Nan Jiang, Email: jiangn678@163.com

Abstract

This research paper addresses the hypothesis that Septin6 is a key regulatory factor influencing amino acid (AA)-mediated cell growth and casein synthesis in dairy cow mammary epithelial cells (DCMECs). DCMECs were treated with absence of AA (AA−), restricted concentrations of AA (AAr) or normal concentrations of AA (AA+) for 24 h. Cell growth, expression of CSN2 and Septin6 were increased in response to AA supply. Overexpressing or inhibiting Septin6 demonstrated that cell growth, expression of CSN2, mTOR, p-mTOR, S6K1 and p-S6K1 were up-regulated by Septin6. Furthermore, overexpressing or inhibiting mTOR demonstrated that the increase in cell growth and expression of CSN2 in response to Septin6 overexpression were inhibited by mTOR inhibition, and vice versa. Our hypothesis was supported; we were able to show that Septin6 is an important positive factor for cell growth and casein synthesis, it up-regulates AA-mediated cell growth and casein synthesis through activating mTORC1 pathway in DCMECs.

Type
Research Article
Copyright
Copyright © Hannah Dairy Research Foundation 2019 

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References

Appuhamy, JA, Knoebel, NA, Nayananjalie, WA, Escobar, J and Hanigan, MD (2012) Isoleucine and leucine independently regulate mTOR signaling and protein synthesis in MAC-T cells and bovine mammary tissue slices. Journal of Nutrition 142, 484491.Google Scholar
Appuhamy, JADRN, Nayananjalie, WA, England, EM, Gerrard, DE, Akers, RM and Hanigan, MD (2014) Effects of AMP-activated protein kinase (AMPK) signaling and essential amino acids on mammalian target of rapamycin (mTOR) signaling and protein synthesis rates in mammary cells. Journal Dairy Science 97, 419429.Google Scholar
Arriola Apelo, SI, Singer, LM, Ray, WK, Helm, RF, Lin, XY, McGilliard, ML, St-Pierre, NR and Hanigan, MD (2014) Casein synthesis is independently and additively related to individual essential amino acid supply. Journal Dairy Science 97, 29983005.Google Scholar
Burgos, SA, Dai, M and Cant, JP (2010) Nutrient availability and lactogenic hormones regulate mammary protein synthesis through the mammalian target of rapamycin signaling pathway. Journal Dairy Science 93, 153161.Google Scholar
Castro Marquez, JJ, Arriola Apelo, SI, Appuhamy, JA and Hanigan, MD (2016) Development of a model describing regulation of casein synthesis by the mammalian target of rapamycin (mTOR) signaling pathway in response to insulin, amino acids, and acetate. Journal Dairy Science 99, 67146736.Google Scholar
Estey, MP, Di Ciano-Oliveira, C, Froese, CD, Bejide, MT and Trimble, WS (2010) Distinct roles of septins in cytokinesis: SEPT9 mediates midbody abscission. Journal of Cell Biology 191, 741749.Google Scholar
Fingar, DC, Richardson, CJ, Tee, AR, Cheatham, L, Tsou, C and Blenis, J (2014) mTOR controls cell cycle progression through its cell growth effectors S6K1 and 4E-BP1/eukaryotic translation initiation factor 4E. Molecular and Cellular Biology 24, 200216.Google Scholar
Gordon, BS, Kazi, AA, Coleman, CS, Dennis, MD, Chau, V, Jefferson, LS and Kimball, SR (2014) Rhoa modulates signaling through the mechanistic target of rapamycin complex 1 (mTORC1) in mammalian cells. Cellular Signalling 26, 461467.Google Scholar
Jiang, N, Wang, Y, Yu, Z, Hu, L, Liu, C, Gao, X and Zheng, S (2015a) WISP3 (CCN6) regulates milk protein synthesis and cell growth through mTOR signaling in dairy cow mammary epithelial cells. DNA and Cell Biology 34, 524533.Google Scholar
Jiang, N, Hu, L, Liu, C, Gao, X and Zheng, S (2015b) 60S ribosomal protein L35 regulates β-casein translational elongation and secretion in bovine mammary epithelial cells. Archives of Biochemistry and Biophysic 583, 130139.Google Scholar
Kaplan, C, Steinmann, M, Zapiorkowska, NA and Ewers, H (2017) Functional redundancy of septin homologs in dendritic branching. Front Cell Developmental Biology 5, 11.Google Scholar
Khudhair, N, Luo, C, Khalid, A, Zhang, L, Zhang, S, Ao, J, Li, Q and Gao, X (2015) 14-3-3γ affects mTOR pathway and regulates lactogenesis in dairy cow mammary epithelial cells. In Vitro Cellular & Developmental Biology-Animal 51, 697704.Google Scholar
Kim, MS, Froese, CD, Xie, H and Trimble, WS (2012) Uncovering principles that control septin-septin interactions. Journal of Biological Chemistry 287, 3040630413.Google Scholar
Kim, SG, Buel, GR and Blenis, J (2013) Nutrient regulation of the mTOR complex 1 signaling pathway. Molecules and Cells 35, 463473.Google Scholar
Li, HM, Wang, CM, Li, QZ and Gao, XJ (2012) MiR-15a decreases bovine mammary epithelial cell viability and lactation and regulates growth hormone receptor expression. Molecules 17, 1203712048.Google Scholar
Lu, LM, Gao, XJ, Li, QZ, Huang, JG, Liu, R and Li, HM (2012) Comparative phosphoproteomics analysis of the effects of L-methionine on dairy cow mammary epithelial cells. Canadian Journal of Animal Science 92, 433442.Google Scholar
Luo, CC, Yin, DY, Gao, XJ, Li, QZ and Zhang, L (2013) Goat mammary gland expression of Cecropin B to inhibit bacterial pathogens causing mastitis. Animal Biotechnology 24, 6678.Google Scholar
Luo, C, Zhao, S, Zhang, M, Gao, Y, Wang, J, Hanigan, MD and Zheng, N (2018) SESN2 negatively regulates cell proliferation and casein synthesis by inhibition the amino acid-mediated mTORC1 pathway in cow mammary epithelial cells. Scientific Reports 8, 3912.Google Scholar
McQuilken, M, Jentzsch, MS, Verma, A, Mehta, SB, Oldenbourg, R and Gladfelter, AS (2017) Analysis of septin reorganization at cytokinesis using polarized fluorescence microscopy. Front Cell Developmental Biology 5, 42.Google Scholar
Mostowy, S and Cossart, P (2012) Septins: the fourth component of the cytoskeleton. Nature Reviews Molecular Cell Biology 13, 183194.Google Scholar
Neubauer, K and Zieger, B (2017) The mammalian septin interactome. Front Cell Developmental Biology 7, 3.Google Scholar
Saxton, RA and Sabatini, DM (2017) mTOR signaling in growth, metabolism, and disease. Cell 169, 361371.Google Scholar
Senger, K, Marka, G, Soller, K, Sakk, V, Florian, MC and Geiger, H (2017) Septin 6 regulates engraftment and lymphoid differentiation potential of murine long-term hematopoietic stem cells. Experimental Hematology 55, 4555.Google Scholar
Spiliotis, ET and Gladfelter, AS (2012) Spatial guidance of cell asymmetry: septin GTPases show the way. Traffic 13, 195203.Google Scholar
Tong, HL, Gao, XJ, Li, QZ, Liu, J, Li, N and Wan, ZY (2011) Metabolic regulation of mammary gland epithelial cells of dairy cow by galactopoietic compound isolated from Vaccariae segetalis. Agricultural Sciences in China 10, 11061116.Google Scholar
Weirich, CS, Erzberger, JP and Barral, Y (2008) The septin family of GTPases: architecture and dynamics. Nature Reviews Molecular Cell Biology 9, 478489.Google Scholar
Wloka, C, Nishihama, R, Onishi, M, Oh, Y, Hanna, J, Pringle, JR, Krauss, M and Bi, E (2011) Evidence that a septin diffusion barrier is dispensable for cytokinesis in budding yeast. Biological Chemistry 392, 813829.Google Scholar
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