Identification of PRG1, a novel progestin-responsive gene with sequence homology to 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.
To define early molecular targets of progestin action, the differential display technique was used to identify genes with altered levels of expression in T-47D breast cancer cells treated with the synthetic progestin ORG 2058 for 3 h. PRG1 was first isolated as a 200-bp cDNA clone and its progestin regulation confirmed by Northern analysis. Cloning of the complete coding region of PRG1 revealed that it shared a high degree of amino acid sequence identity with isoforms of the enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase from several tissues and species. Expression of PRG1 mRNA was observed in several normal breast epithelial and breast cancer cell lines and in a variety of human tissues, with highest expression in the breast, aorta, and brain. In T-47D cells, PRG1 mRNA was rapidly and transiently induced by progestins, expression peaking between 2 and 4 h and returning to control levels by 12 h. Progestin-induced increases in PRG1 mRNA were inhibited by the progestin antagonist RU 486 and occurred via the progesterone receptor. Progestin induction of PRG1 mRNA was also inhibited by actinomycin D but not by cycloheximide. PRG1 is therefore a novel human gene that is directly regulated by progestins via the progesterone receptor.
Hamilton
J A
JA
Garvan Institute of Medical Research, St. Vincent's Hospital, Sydney, New South Wales, Australia.
Callaghan
M J
MJ
Sutherland
R L
RL
Watts
C K
CK
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001943:Breast Neoplasms / Q000201:enzymology / Q000235:genetics; D002417:Cattle; D002453:Cell Cycle / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D006597:Fructose-Bisphosphatase / Q000737:chemistry* / Q000235:genetics; D005786:Gene Expression Regulation; D006019:Glycolysis / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010732:Phosphofructokinase-1 / Q000737:chemistry* / Q000235:genetics; D025481:Phosphofructokinase-2; D011282:Pregnenediones / Q000494:pharmacology; D045167:Progesterone Congeners / Q000494:pharmacology; D011372:Progestins / Q000502:physiology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011506:Proteins / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000302:isolation & purification; D016415:Sequence Alignment; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D011282:Pregnenediones; D045167:Progesterone Congeners; D011372:Progestins; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D012333:RNA, Messenger; C020175:16 alpha-ethyl-21-hydroxy-19-nor-4-pregnene-3,20-dione; C107345:PFKFB3 protein, human; D025481:Phosphofructokinase-2; D010732:Phosphofructokinase-1; D006597:Fructose-Bisphosphatase