Prolactin-dependent growth and gamma-casein gene expression in Ba/F3 cells transfected with a long form of mouse mammary prolactin receptor.
Complementary DNA (cDNA) encoding a long form of prolactin receptor (PRL-RL) was cloned from mouse mammary gland by PCR using primers designed from the noncoding regions of previously reported rat ovarian PRL-RL cDNA. The nucleotide sequence encoding the extracellular and transmembrane domains of PRL-RL is completely identical to that of short forms of mouse PRL-R. The amino acid sequence deduced from the nucleotide sequence of mouse PRL-RL is 91% identical to that of rat PRL-RL. To address the question of whether or not the cloned mouse PRL-RL cDNA encodes a functional PRL-RL we transfected Ba/F3 IL-3-dependent murine pro-B lymphoid cells with the cDNA. By culturing the transfected cells in a medium which contained PRL in place of IL-3, we selected 5 PRL-dependent clones. All of these PRL-dependent clones, BaF/PD cells, expressed PRL-RL mRNA. In addition, BaF/PD cells expressed mammary-specific gamma-casein mRNA in response to PRL and dexamethasone. Based on these results, it was concluded that the mouse mammary PRL-RL cDNA cloned in this study is functionally active in mediating both PRL-dependent growth and mammary-specific gene expression.
Sasaki
M
M
Research Laboratory, Zenyaku Kogyo Co., Ltd., Tokyo, Japan.
Sasaki
T
T
Enami
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001402:B-Lymphocytes / Q000378:metabolism; D001483:Base Sequence; D002364:Caseins / Q000235:genetics*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D015870:Gene Expression; D008321:Mammary Glands, Animal / Q000737:chemistry*; D051379:Mice; D008969:Molecular Sequence Data; D011388:Prolactin / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011981:Receptors, Prolactin / Q000737:chemistry / Q000235:genetics*; D014162:Transfection
D002364:Caseins; D018076:DNA, Complementary; D012333:RNA, Messenger; D011981:Receptors, Prolactin; D011388:Prolactin