Cloning of a protein binding to the most proximal Pit-1 binding element of prolactin gene from human pituitary cDNA library.
A human pituitary cDNA library was screened using a yeast one-hybrid system to find a factor binding Pit-1 binding elements in the PRL gene other than Pit-1. Beside colonies containing Pit-1 or Oct-1 cDNA, three colonies contained mPOU cDNA, a member of the POU protein family. Immunohistochemical analysis showed mPOU-like immunoreactivity was present in human PRL-producing pituitary tumors but not in non-functioning pituitary tumors. Mobility shift analysis revealed that mPOU bound to Pit-1 binding elements of the PRL gene, 1P and 3P. mPOU activated the expression of 0.6 k PRL and 7x1P reporter genes in the presence of Pit-1 and cAMP, although it did not enhance Pit-1-induced expression of 7x3P reporter gene. These findings suggest that mPOU is involved in the activation of the PRL gene by cAMP through 1P in the presence of Pit-1.
Fumoto
Mariko
M
Division of Endocrinology/Metabolism, Neurology and Hematology/Oncology, Department of Clinical Molecular Medicine, Kobe University Graduate School of Medicine, Kobe, Japan.
Okimura
Yasuhiko
Y
Sakagami
Yoshio
Y
Iguchi
Genzo
G
Kishimoto
Masahiko
M
Takahashi
Yutaka
Y
Kaji
Hidesuke
H
Chihara
Kazuo
K
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D019556:COS Cells; D002118:Calcium / Q000378:metabolism; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D000242:Cyclic AMP / Q000031:analogs & derivatives* / Q000494:pharmacology; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation / Q000187:drug effects; D015723:Gene Library; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D051863:Host Cell Factor C1; D006801:Humans; D007150:Immunohistochemistry; D009154:Mutation; D050811:Octamer Transcription Factor-1; D050809:POU Domain Factors; D010902:Pituitary Gland / Q000378:metabolism*; D011388:Prolactin / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013873:Thionucleotides / Q000494:pharmacology; D050817:Transcription Factor Pit-1; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D018076:DNA, Complementary; D004268:DNA-Binding Proteins; C497692:HCFC1 protein, human; D051863:Host Cell Factor C1; D050811:Octamer Transcription Factor-1; D050809:POU Domain Factors; C494187:POU1F1 protein, human; C494165:POU2F1 protein, human; D011993:Recombinant Fusion Proteins; D013873:Thionucleotides; D050817:Transcription Factor Pit-1; D014157:Transcription Factors; C015929:8-((4-chlorophenyl)thio)cyclic-3',5'-AMP; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D011388:Prolactin; D000242:Cyclic AMP; D002118:Calcium