title: Prolactin-regulated pim-1 transcription: identification of critical promoter elements and Akt signaling.
abstract: Previously we showed that the distal element (DE) (-427 to -336 bp) within the pim-1 promoter appeared to regulate its prolactin (PRL)-induced transcription. To determine which specific DE sequences conferred PRL responsiveness, seven 12-bp deletion mutants ligated upstream of the chloramphenicol acetyltransferase gene were transfected into FDC/Nb2 cells. Results from promoter/ reporter studies showed that sequential 12-bp deletions of the DE significantly (p < 0.001) reduced PRL responsiveness. An additional site, nuclear factor-1 (-224 to -217), was also mutated by deletion or point mutation; both abrogated promoter activation by PRL (p < 0.0001). In other experiments, PRL signaling to pim-1 expression was investigated in FDC/Nb2 cells stably expressing the wild-type (WT) Jak2 cDNA or a carboxy-terminal kinase-deficient Jak2 mutant and in cells infected with adenoviral constructs of WT-Akt or dominant negative Akt. Altered Jak2 did not affect PRL-stimulated pim-1 expression while inhibition of Akt attenuated its transcription. We conclude that the DE and NF-1 half-site mediate PRL responsiveness of the pim-1 promoter. Moreover, the accumulated evidence does not support a role for the Jak2/Stat signaling pathway but, instead, implicates that Akt activation was a component of PRL-induced pim-1 transcription.
authors.affiliation: College of Pharmacy, University of Cincinnati Medical Center, Cincinnati, OH 45267-0004, USA.
authors.forename: Nithya
authors.initials: N
authors.lastname: Krishnan
authors.forename: Huiqi
authors.initials: H
authors.lastname: Pan
authors.forename: Donna J
authors.initials: DJ
authors.lastname: Buckley
authors.forename: Arthur
authors.initials: A
authors.lastname: Buckley
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D053614:Janus Kinase 2; D008223:Lymphoma; D011388:Prolactin / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt; D051573:Proto-Oncogene Proteins c-pim-1; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015398:Signal Transduction / Q000502:physiology; D014158:Transcription, Genetic / Q000502:physiology
chemical_list: D011518:Proto-Oncogene Proteins; D012333:RNA, Messenger; D011388:Prolactin; D011505:Protein-Tyrosine Kinases; C507927:Jak2 protein, rat; D053614:Janus Kinase 2; C494920:Akt1 protein, rat; C496637:Pim1 protein, rat; D017346:Protein Serine-Threonine Kinases; D051057:Proto-Oncogene Proteins c-akt; D051573:Proto-Oncogene Proteins c-pim-1