Ratio of lysine to methionine alters expression of genes involved in milk protein transcription and translation and mTOR phosphorylation in bovine mammary cells

Xuemei Nan, Dengpan Bu, Xiyan Li, Jiaqi Wang, Hongyang Wei, Han Hu, Lingyun Zhou, Juan J. Loor

Research output: Contribution to journalArticle

Abstract

This study was conducted to determine the optimum ratio of lysine and methionine (Lys:Met) to enhance milk protein concentration in vitro, focusing on the regulation of genes related to the JAK2-STAT5 and the mammalian target of rapamycin (mTOR) signaling pathways. A preliminary dose response study revealed that casein concentration peaked (2.5-2.7 ppm) at a supplemental Lys concentration of 1.2 mM and Met at 0.5 mM. At the peak casein concentration cell proliferation rate also was higher. Furthermore, the expression of CSN1S1, CSN1S2, CSN2, CSN3, LALBA, JAK2, STAT5, and MTOR was upregulated with both Lys and Met compared with the control. A subsequent experiment was conducted as a 5 × 3 factorial design with supplemental Lys plus Met at different ratios. When the supplemental concentration of Lys was 1.2 mM and Met was 0.4 mM (~3:1), the concentration of casein peaked. Therefore, we measured gene expression, mTOR protein expression, and phosphorylated mTOR (p-mTOR) in cultures incubated with 3:1 Lys:Met (Lys&Met). Expression of CSN1S1 and LALBA were the most highly expressed genes (P < 0.01). The upregulation of CSN2, CSN3, CSN1S2 isoforms (P < 0.01) and JAK2, ELF5, mTOR (P < 0.05) was also observed. Total mTOR protein expression was greater (P < 0.05) with Lys alone and also Lys&Met. However, Lys&Met resulted in the greatest (P < 0.05) p-mTOR. Results suggest that peak concentration of casein at a supplemental 3:1 Lys:Met is driven in part via upregulation of the mRNA expression of components of the JAK-STAT and mTOR pathways.

Original languageEnglish (US)
Pages (from-to)268-275
Number of pages8
JournalPhysiological genomics
Volume46
Issue number7
DOIs
StatePublished - Apr 1 2014

Keywords

  • Amino acids
  • Mammary cell
  • Milk protein

ASJC Scopus subject areas

  • Physiology
  • Genetics

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