Reconstitution of the protein kinase A response of the rat prolactin promoter: differential effects of distinct Pit-1 isoforms and functional interaction with Oct-1.
PRL gene transcription is primarily regulated by dopamine, which lowers cAMP levels and inhibits protein kinase A (PKA) activity. Current data indicate that the cAMP/PKA response maps to the most proximal Pit-1/Pit-1beta binding site footprint I (FP I) on the rat PRL (rPRL) promoter. Pit-1, a POU-homeo domain transcription factor, is specifically expressed in the anterior pituitary and is required both for the normal development of anterior pituitary cell types, somatotrophs, lactotrophs, and thyrotrophs, and for the expression of their hormones: GH, PRL, and TSHbeta. Pit-1 has been shown to functionally interact, via FP I, with several transcription factors, including Oct-1, a ubiquitous homeobox protein, and thyrotroph embryonic factor, which is found in lactotrophs, to activate basal rPRL promoter activity. Pit-1beta/GHF-2, a distinct splice isoform of Pit-1, acts to inhibit Ras-activated transcription from the rPRL promoter, which is mediated by a functional interaction between Pit-1 and Ets-1 at the most distal Pit-1 binding site (FP IV). In this manuscript we show 1) that the Pit-1beta isoform not only fails to block PKA activation, but is, in fact, a superior mediator of the PKA response; 2) that the PKA response requires intact POU-specific and POU-homeo domains of Pit-1; and 3) that Oct-1, but not thyrotroph embryonic factor, functions as a Pit-1-interacting factor to mediate an optimal PKA response.
Diamond
S E
SE
Department of Medicine, Colorado Cancer Center, University of Colorado Health Sciences Center, Denver 80262, USA.
Chiono
M
M
Gutierrez-Hartmann
A
A
D000818:Animals; D050976:Basic-Leucine Zipper Transcription Factors; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000737:chemistry; D006367:HeLa Cells; D051863:Host Cell Factor C1; D006801:Humans; D008156:Luciferases / Q000032:analysis; D008163:Luminescent Measurements; D016296:Mutagenesis; D050811:Octamer Transcription Factor-1; D010957:Plasmids / Q000737:chemistry; D016133:Polymerase Chain Reaction; D011388:Prolactin / Q000235:genetics / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000502:physiology; D020033:Protein Isoforms; D051381:Rats; D015398:Signal Transduction; D050817:Transcription Factor Pit-1; D014157:Transcription Factors / Q000502:physiology*; D014162:Transfection