The transcriptional effect of WT1 is modulated by choice of expression vector.
The WT1 Wilms' tumor suppressor gene encodes a zinc finger transcription factor which plays a critical role in renal and genitourinary development. The WT1 protein was reported to both activate and repress transcription. We found that the transcriptional effect of WT1 on the Egr1 promoter could be modulated by the use of expression vectors containing different promoters. WT1 activated the Egr1 promoter when expression of WT1 was driven by the Rous sarcoma virus promoter. In contrast, a cytomegalovirus (CMV) promoter-containing WT1 expression vector repressed the Egr1 promoter. However, WT1 activated transcription of a simple test promoter, EGR3tkCAT, regardless of the expression vector used. Co-transfection of the parental CMV-based vector strongly depressed the basal activity of the Egr1-CAT reporter, suggesting that the CMV promoter competes with the Egr1 promoter for transcription factors or co-factors which may be required for activation by WT1. In support of this hypothesis, WT1 was converted from an activator to a repressor by co-transfection of an excess of the parental CMV-based vector. These results provide an important caveat to the interpretation of co-transfection studies and confirm the bi-functional nature of the WT1 transcription factor.
Reddy
J C
JC
Brookdale Center for Molecular Biology, Mount Sinai School of Medicine, New York, New York 10029, USA.
Hosono
S
S
Licht
J D
JD
D000818:Animals; D001358:Avian Sarcoma Viruses; D002460:Cell Line; D002522:Chlorocebus aethiops; D003587:Cytomegalovirus; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000378:metabolism*; D015870:Gene Expression; D016162:Genes, Wilms Tumor; D005822:Genetic Vectors; D007668:Kidney; D007700:Kinetics; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid; D014157:Transcription Factors / Q000096:biosynthesis* / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection; D025721:WT1 Proteins; D016335:Zinc Fingers
D004268:DNA-Binding Proteins; D014157:Transcription Factors; D025721:WT1 Proteins