title: Positive and negative regulatory elements are involved in transcriptional control of the rat glucokinase gene in the insulin producing cell line HIT M2.2.2.
abstract: Nested deletion mutants of the 5' flanking region of the beta-cell transcription unit of the rat glucokinase gene (r beta GK) were fused to the CAT-reporter gene. Transient expression studies in HIT M2.2.2 and BHK21 cells revealed a distal (upstream of -359) and a proximal promoter region (between -278/-49) harbouring positive and negative regulatory elements. DNaseI footprinting revealed three protected areas between nucleotides -190 and -60. DNA-elements playing a crucial role in transcriptional control of the insulin genes (IEB- and CT-motifs) have been detected within the proximal promoter region and contribute to beta-cell specific gene regulation. 3' deletion analysis revealed that DNA-elements located downstream from transcription initiation sites (up to +123) contribute to transcriptional regulation.
authors.affiliation: Institut für Biochemie, Klinikum, Medizinische Fakultät, Ernst-Moritz-Arndt-Universität Greifswald, Germany.
authors.forename: I B
authors.initials: IB
authors.lastname: Leibiger
authors.forename: R
authors.initials: R
authors.lastname: Walther
authors.forename: U
authors.initials: U
authors.lastname: Pett
authors.forename: B
authors.initials: B
authors.lastname: Leibiger
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis / Q000378:metabolism; D006224:Cricetinae; D017931:DNA Primers; D003850:Deoxyribonuclease I; D015870:Gene Expression; D005941:Glucokinase / Q000096:biosynthesis* / Q000235:genetics*; D007328:Insulin / Q000096:biosynthesis; D007515:Islets of Langerhans / Q000378:metabolism*; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D051381:Rats / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid; D017384:Sequence Deletion; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D017931:DNA Primers; D007328:Insulin; D015500:Chloramphenicol O-Acetyltransferase; D005941:Glucokinase; D003850:Deoxyribonuclease I