A proportion of secreted pituitary prolactin (PRL) is phosphorylated. However, because most commercial sources of PRL are recombinant proteins without posttranslational modification, the importance of PRL phosphorylation to the production of milk proteins is an understudied area. Here, we have examined the effect of PRL phosphorylation on expression of the milk protein, β-casein, using a phospho-stable mimic of the phosphorylated form (S179D-PRL) and analyzing promoter activation and mRNA stability over a 7-day treatment period in response to this and unmodified PRL. At equivalent concentrations, the phospho-mimic showed a superior ability to activate a −2300 → +490 region of the promoter, but not an artificial promoter consisting of three Stat5 consensus sites upstream of a minimal promoter. Unlike unmodified PRL, S179D-PRL was also able to stabilize β-casein mRNA. These effects of S179D-PRL were eliminated by incubation in the MAP kinase pathway inhibitor, U0126, bringing promoter activation down to the level seen with unmodified PRL and essentially eliminating the effect on mRNA stability. These results support an important role for the posttranslational phosphorylation of PRL and signaling through the MAP kinase pathway in the production of this milk protein.
Part of the book: Milk Protein