Promoter suppression in cultured mammalian cells can be blocked by the chicken beta-globin chromatin insulator 5'HS4 and matrix/scaffold attachment regions.
Studies have indicated that two transcriptionally active units can repress one another when they lie adjacent in head-to-tail tandem on a chromosome. Repression of a downstream (3') unit by an upstream (5') unit is known as "transcriptional interference", whereas repression of a 5' unit by a 3' unit is termed "promoter suppression". These two processes can occur between head-to-tail tandem copies of a transgene, or between transgenes and adjacent chromosomal genes. Interference can be blocked by inserting a transcription terminator between adjacent units. Here, we report that "promoter suppression" could be blocked by the insulator 5' DNaseI hypersensitive site 4, or matrix/scaffold attachment regions (MAR/SARs), when these elements were interposed between adjacent units. Because intergenic spacers of many repeated eukaryotic genes contain MAR/SARs and insulators, our observations suggest that these elements have the ability to segregate repeated genes into domains that act independently of one another. Our observations also suggest strategies to design transgenes that can act as autonomous units of expression.
Villemure
J F
JF
Molecular Biology Program and Department of Microbiology and Immunology, Université de Montréal, Montréal, Québec, Canada.
Savard
N
N
Belmaaza
A
A
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002645:Chickens / Q000235:genetics*; D002843:Chromatin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004247:DNA / Q000235:genetics / Q000378:metabolism; D020868:Gene Silencing; D017930:Genes, Reporter / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics; D005914:Globins / Q000235:genetics*; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D018139:Simplexvirus / Q000235:genetics; D013728:Terminator Regions, Genetic / Q000235:genetics; D013937:Thymidine Kinase / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics; D014162:Transfection
D002843:Chromatin; D056572:Histone Deacetylase Inhibitors; D005914:Globins; D004247:DNA; D013937:Thymidine Kinase; D006655:Histone Deacetylases