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Variation in the coding and 3′ untranslated regions of the porcine prolactin receptor short form modifies protein expression and function
Author(s) -
Trott Josephine F.,
Freking Bradley A.,
Hovey Russell C.
Publication year - 2014
Publication title -
animal genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.756
H-Index - 81
eISSN - 1365-2052
pISSN - 0268-9146
DOI - 10.1111/age.12100
Subject(s) - biology , haplotype , untranslated region , single nucleotide polymorphism , coding region , gene isoform , microbiology and biotechnology , prolactin receptor , exon , genetics , prolactin , messenger rna , gene , genotype , endocrinology , hormone
Summary The actions of prolactin ( PRL ) are mediated by both long ( LF ) and short isoforms ( SF ) of the PRL receptor ( PRLR ). Here, we report on a genetic and functional analysis of the porcine PRLR ( pPRLR ) SF. Three single nucleotide polymorphisms ( SNP s) within exon 11 of the p PRLR ‐ SF give rise to four amino acid haplotypes of the intracellular domain. We identified the dimorphic insertion of a short interspersed repetitive DNA element ( PRE ‐1) along with 32 SNP s and four other insertion/deletion sites within the 3′ untranslated region (UTR) of p PRLR ‐ SF . The PRE ‐1 element reduced protein translation in vitro by 75%, whereas the combination of 10 SNP s and one insertion/deletion decreased translation by 50%. Full‐length cDNA s for all four haplotypes of p PRLR ‐ SF were cloned behind the elongation factor 1‐alpha promoter and functionally analyzed in vitro . None of the haplotypes could initiate transcription from the ß‐casein promoter, whereas all four were dominant negatives against PRL ‐activation of the pPRLR ‐LF. Two of the haplotypes completely inhibited pPRLR ‐LF activity at a four‐fold excess, whereas the others required a six‐fold excess to impart the same effect. The ligand binding affinities of the pPRLR ‐SF haplotypes did not differ. Expression of the p PRLR ‐SF increased linearly during gestation in the endometrium and was hormonally regulated in a tissue‐specific manner in the mammary glands and uterus. In conclusion, we identified a PRE‐1 and other SNPs in the p PRLR ‐SF 3′ UTR that reduce protein expression and four haplotypes of the pPRLR ‐SF that suppress pPRLR ‐LF signaling and may differentially impact the phenotypic effects of PRL in vivo .