Nucleic Acids Research, 1994, Vol. 22, No. 21 4386-4394
© 1994
MOLECULAR BIOLOGY |
SARs stimulate but do not confer position independent gene expression
Departments of Molecular Biology and Biochemistry, University of Geneva 30 Quai Ernest-Ansermet, CH 1211 Geneva 4, Switzerland
*To whom correspondence should be addressed at; Laboratoire de Biologie Moléculaire Eucaryote, CNRS UPR 9006, 118 route de Narbonne, 31062 Toulous, France
Received July 28, 1994. Revised September 20, 1994. Accepted September 20, 1994.
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 ( {small tilde}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.
+Present address: Department of Animal Biology, University of Geneva, 154 route de Malagnou, 1224 Chêne-Bougeries
Department of Cellular Biology, university of Geneva, 30, Quai Ernest-Ansermet, CH 1211 Geneva 4, Switzerland
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