Characterization of the 5'-flanking region of the human preprogalanin gene.
Expression of the human galanin gene was analysed using a 3.5-kb DNA fragment comprising the 5'-flanking sequence of the gene. This sequence contains a TATA box (ATATATA) preceded by numerous potential binding sites for transcription factors such as SP1, AP2, and NF kappa B. Three half-palindromic estrogen response elements (EREs, GGTCA) are also found at positions -1,162, -361, and -122 bp relative to the transcription start site. To localize functionally important portions of the promoter region, several shorter fragments of the galanin 5'-flanking region were placed upstream from the chloramphenicol acetyltransferase (CAT) reporter gene. In transient transfection assays, all constructs demonstrated substantial transcriptional activity in both rat glioma/mouse neuroblastoma hybrid cells (NG108-15) and Chinese hamster ovary (CHO-K1) cells. Comparison of the basal expression levels of the different constructs suggests the presence of a negative modulator between positions -1,891 and -207. When cotransfected into NG108-15 cells with the human estrogen receptor cDNA, estrogen did not induce transcription of the human galanin gene at physiological levels of estrogen receptor, although transcription was induced up to 30-fold in the presence of high levels of receptor.
Kofler
B
B
Garvan Institute of Medical Research, St. Vincent's Hospital, Darlinghurst, NSW, Australia.
Evans
H F
HF
Liu
M L
ML
Falls
V
V
Iismaa
T P
TP
Shine
J
J
Herzog
H
H
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D004247:DNA / Q000378:metabolism; D004967:Estrogens / Q000502:physiology; D019004:Galanin; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D006801:Humans; D008969:Molecular Sequence Data; D010455:Peptides / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011498:Protein Precursors / Q000235:genetics; D011960:Receptors, Estrogen / Q000378:metabolism; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured
D004967:Estrogens; C056567:GAL protein, human; C489169:Gal protein, rat; D010455:Peptides; D011498:Protein Precursors; D011960:Receptors, Estrogen; D019004:Galanin; D004247:DNA