TY - JOUR
T1 - Integration of tetracycline regulation into a cell-specific transcriptional enhancer
AU - Rose, Scott D.
AU - MacDonald, Raymond J.
PY - 1997/2/21
Y1 - 1997/2/21
N2 - The pancreas-specific transcriptional enhancer of the rat elastase I gene was modified by substituting, in turn, each of its three individual constitutive elements with the tetO element, which confers regulation by exogenous tetracycline in the presence of the hybrid tetO binding transactivator (tTA). Whereas the unmodified enhancer was active in transfected acinar tumor cells, substitution of individual elements with the tet-responsive element abolished activity. The modified enhancers were reactivated in the presence of the tTA and, upon addition of tetracycline, were silenced. Thus, substitution of individual enhancer elements renders the enhancer responsive to regulation by tetracycline. Moreover, the tTA- activated levels were 2-8-fold greater than the unmodified enhancer. The acinar cell specificity of the unmodified enhancer was retained; none of the tetO-substituted enhancers were activated by tTA in a variety of nonacinar cell lines. These results show that a foreign and artificial transcriptional activator, tTA, can be incorporated into an enhancer to create a novel, efficient, and regulatable transcriptional control region whose cell specificity is retained.
AB - The pancreas-specific transcriptional enhancer of the rat elastase I gene was modified by substituting, in turn, each of its three individual constitutive elements with the tetO element, which confers regulation by exogenous tetracycline in the presence of the hybrid tetO binding transactivator (tTA). Whereas the unmodified enhancer was active in transfected acinar tumor cells, substitution of individual elements with the tet-responsive element abolished activity. The modified enhancers were reactivated in the presence of the tTA and, upon addition of tetracycline, were silenced. Thus, substitution of individual enhancer elements renders the enhancer responsive to regulation by tetracycline. Moreover, the tTA- activated levels were 2-8-fold greater than the unmodified enhancer. The acinar cell specificity of the unmodified enhancer was retained; none of the tetO-substituted enhancers were activated by tTA in a variety of nonacinar cell lines. These results show that a foreign and artificial transcriptional activator, tTA, can be incorporated into an enhancer to create a novel, efficient, and regulatable transcriptional control region whose cell specificity is retained.
UR - http://www.scopus.com/inward/record.url?scp=0031051518&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0031051518&partnerID=8YFLogxK
U2 - 10.1074/jbc.272.8.4735
DO - 10.1074/jbc.272.8.4735
M3 - Article
C2 - 9030525
AN - SCOPUS:0031051518
SN - 0021-9258
VL - 272
SP - 4735
EP - 4739
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 8
ER -