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.
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.
Leibiger
I B
IB
Institut für Biochemie, Klinikum, Medizinische Fakultät, Ernst-Moritz-Arndt-Universität Greifswald, Germany.
Walther
R
R
Pett
U
U
Leibiger
B
B
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
D017931:DNA Primers; D007328:Insulin; D015500:Chloramphenicol O-Acetyltransferase; D005941:Glucokinase; D003850:Deoxyribonuclease I