The prolactin receptor from the brushtail possum (Trichosurus vulpecula): cDNA cloning, expression and functional analysis |
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Authors: | Demmer J |
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Affiliation: | Dairy Science Group, AgResearch, Ruakura Research Centre, Hamilton, New Zealand. demmerj@agresearch.cri.nz |
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Abstract: | A full length, prolactin receptor cDNA clone has been isolated from the brushtail possum (Trichosurus vulpecula). This clone encodes a 625 amino acid protein which shares 60-70 and 54% sequence identity with prolactin receptor (long form) sequences from mammalian and avian species, respectively. Sequence similarity was highest in the extra-cellular, hormone-binding domain and in specific regions of the intracellular domain which regulates prolactin receptor signalling in cells. Prolactin receptor mRNA was detected in a wide range of possum tissues and in the mammary gland the PRL-R gene was differentially expressed during lactation with peak mRNA levels being detected during the first 6 days of lactation and after day 115 throughout late lactation. This pattern of PRL-R mRNA expression in the mammary gland is similar to that observed for circulating prolactin in the lactating possum. In CHO cells transiently transfected with the possum prolactin receptor, expression of a beta-lactoglobulin promoter/reporter gene construct was increased 3-fold by adding prolactin. The possum prolactin receptor is therefore capable of binding ovine prolactin and activating the Jak2/Stat5 signalling cascade. This provides evidence for the highly conserved nature of the prolactin signalling pathway in mammalian evolution. |
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