Cloning and functional characterization of allelic variation in the porcine prolactin receptor.
Prolactin (PRL) regulates various functions in pigs including reproduction, mammary development and lactation. We used 5'-rapid amplification of cDNA ends (5'-RACE) to clone three full-length alleles of the porcine PRL receptor (pPRLR) from Landrace (alleles LR2 and LR4) and Yucatan miniature (MP) pigs, corresponding to the A and B alleles previously reported to be associated with reproductive traits. When expressed in Chinese hamster ovary (CHO-K1) cells, all three pPRLRs transduced differentiation signals to a beta-casein promoter with the same effectiveness, where human growth hormone (hGH) and porcine PRL (pPRL) were more effective ligands than ovine PRL (oPRL). The pPRLR had a lower binding affinity for oPRL than pPRL while binding affinity for hGH was not different between the three pPRLR variants. The pPRLRs primarily localized to the cytoplasm with perinuclear concentration. In conclusion, we have cloned three allelic variants of the pPRLR and have functionally characterized these as different from the hPRLR. However, our data do not support the proposal that allelic variation of the pPRLR confers functional differences in vivo.
Trott
Josephine F
JF
Lactation and Mammary Gland Biology Group, Department of Animal Science, University of Vermont, 221 Terrill Hall, 570 Main Street, Burlington, VT 05405, United States.
Farley
Nicholas R
NR
Taatjes
Douglas J
DJ
Hovey
Russell C
RC
D000483:Alleles; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern / Q000662:veterinary; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D014644:Genetic Variation; D019382:Human Growth Hormone / Q000378:metabolism / Q000494:pharmacology; D007150:Immunohistochemistry; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D011388:Prolactin / Q000378:metabolism / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D012313:RNA / Q000737:chemistry / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique / Q000662:veterinary; D011981:Receptors, Prolactin / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D013552:Swine; D013556:Swine, Miniature / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000662:veterinary
D011981:Receptors, Prolactin; D019382:Human Growth Hormone; D012313:RNA; D011388:Prolactin