Transactivation of pancreas-specific gene sequences in somatic cell hybrids.
Enhancer/promoter elements from two pancreas-specific genes, those encoding amylase and elastase, were ligated to the bacterial GPT gene. The resulting construct can be used to select for expression of gene products which activate these pancreas-specific promoters in hybrid cells. The selectable GPT construct was stably transferred into several cell lines either directly or by cotransfection with pSV2Neo. GPT was expressed when transferred to pancreatic cell lines but not when transferred to GPT-fibroblast (L) cells or hepatoma cells. When the transformed L cells and hepatoma cells were fused with pancreatic cell lines, GPT was activated in the hybrid cells. Endogenous pancreas-specific genes from the L-cell and hepatoma parents were also activated in the hybrids. In addition, a pancreas-specific nuclear protein, PTF1, was produced in pancreatic and hybrid cells, correlating with GPT expression. The transformed L cells and hepatoma cells thus contained a nonexpressed construct which could be activated in trans by factors present in pancreatic cells. The hepatoma hybrid also continued to produce albumin, demonstrating the coexpression of liver and pancreas-specific genes in the hybrid-cell population. Cell lines carrying the amylase/elastase/GPT construct may be useful as a selection system for cloning of pancreatic transcription activators.
Wu
K J
KJ
Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Samuelson
L C
LC
Howard
G
G
Meisler
M H
MH
Darlington
G J
GJ
D000418:Albumins / Q000378:metabolism; D000681:Amylases / Q000235:genetics*; D000818:Animals; D015139:Blotting, Southern; D043424:Carboxypeptidase B; D002268:Carboxypeptidases; D003001:Cloning, Molecular; D004742:Enhancer Elements, Genetic; D006801:Humans; D006822:Hybrid Cells; D008114:Liver Neoplasms, Experimental; D051379:Mice; D009928:Organ Specificity / Q000235:genetics; D010179:Pancreas / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011506:Proteins / Q000378:metabolism; D014158:Transcription, Genetic; D015533:Transcriptional Activation; D014162:Transfection; D014170:Transformation, Genetic; D014407:Tumor Cells, Cultured
D000418:Albumins; C058173:CPB1 protein, human; D011506:Proteins; D000681:Amylases; D002268:Carboxypeptidases; D043424:Carboxypeptidase B; D010196:Pancreatic Elastase