Identification and characterization of the promoter region of the GRM3 gene.
We have recently described the genomic organisation of the human metabotropic glutamate receptor 3 (GRM3) gene. The putative promoter region is characterised by the presence of a CCAAT and Sp1 site and the absence of a TATA box. Using a reporter gene assay, now we describe the functional activity of GRM3 promoter by transient transfection in both human neuroblastoma and astroglioma cell lines. Deletion of the CCAAT box and Sp1 site resulted in a pronounced reduction of reporter gene expression in both cell types, which indicates that these elements to correspond to the core promoter region. Moreover, we found that the genomic sequence 140 bp upstream of the first transcription initiation site appears to contain regulatory promoter elements for a preferential transcription of the gene in neuroblastoma cells. We also provide evidence that the genomic sequence spanning exon I, corresponding to the GRM3 5'-untranslated region, contains a negative regulatory element that represses gene transcription.
Corti
C
C
Cambridge Brain Bank Laboratory, Department of Pathology, University of Cambridge, Level 3 Laboratory Block Addenbrooke's Hospital, Hills Road, Cambridge, CB2 2QQ, United Kingdom. cc18322@gsk.com
Xuereb
J H
JH
Corsi
M
M
Ferraguti
F
F
D020121:5' Untranslated Regions; D001254:Astrocytoma; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D005796:Genes; D017930:Genes, Reporter; D006801:Humans; D008969:Molecular Sequence Data; D009447:Neuroblastoma; D011401:Promoter Regions, Genetic; D018094:Receptors, Metabotropic Glutamate / Q000235:genetics*; D017384:Sequence Deletion
D020121:5' Untranslated Regions; D018094:Receptors, Metabotropic Glutamate; C102718:metabotropic glutamate receptor 3