SARs stimulate but do not confer position independent gene expression.
Two minimal scaffold-associated regions (SARs) from Drosophila were tested in stably transformed cells for their effects on the expression of reporter genes. The expression of genes bounded by two SARs is consistently stimulated by about 20- to 40-fold, if the average of a pool of cell transformants is analyzed. However, analysis of individual, stable cell transformants demonstrates that flanking SAR elements do not confer position-independent expression on the reporter gene and that the extent of position-dependent variegation is similarly large with or without the flanking SAR elements. The SAR stimulation of expression is observed in stable but not in transiently transfected cell lines. The Drosophila scs and scs' boundary elements, which do not bind to the nuclear matrix in vitro, are only about one-tenth as active as SARs in stimulating expression in stable transformants. Interestingly, the SAR stimulatory effect can be blocked by a fragment containing CpG islands (approximately 70% GC), if positioned between the SAR and the enhancer. In contrast, when inserted in the same position, control fragments, such as the scs/scs' elements, do not interfere with SAR function.
Department of Molecular Biology, University of Geneva, Switzerland.
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Poljak
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Seum
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Mattioni
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Laemmli
D000818:Animals; D001482:Base Composition; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics*; D004247:DNA / Q000737:chemistry*; D004330:Drosophila / Q000235:genetics; D004742:Enhancer Elements, Genetic; D015870:Gene Expression; D017930:Genes, Reporter; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D006367:HeLa Cells; D006657:Histones / Q000235:genetics; D006801:Humans; D007739:L Cells / Q000378:metabolism; D051379:Mice; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013539:Simian virus 40 / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection
D018840:HSP70 Heat-Shock Proteins; D006657:Histones; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase