Identification of the cellular protein encoded by the human Wilms' tumor (WT1) gene.
A putative tumor-suppressor gene (wt1) located at chromosome 11p13 and involved in Wilms' tumor development has recently been identified as a zinc finger polypeptide-encoding gene. The purpose of this study was to characterize the protein encoded by the human wt1 gene. The region spanning the entire zinc finger domain was amplified by polymerase chain reaction (PCR) and subcloned in the pATH 3 expression vector. Polyclonal antibodies against the fused TrpE-WT protein were raised. These antibodies immunoprecipitated a 49- to 51-kDa protein from hematopoietic tumor cells labeled in vivo with [35S]methionine. Subcellular fractionation and immunohistochemistry followed by confocal microscopy indicated that the Wilms' tumor gene product (WT1) is mainly localized within the nucleus.
Telerman
A
A
Institute of Interdisciplinary Research, School of Medicine, Free University of Brussels (ULB), Belgium.
Dodemont
H
H
Degraef
C
C
Galand
P
P
Bauwens
S
S
Van Oostveldt
P
P
Amson
R B
RB
D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D002880:Chromosomes, Human, Pair 11; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D004268:DNA-Binding Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics*; D016147:Genes, Tumor Suppressor; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry; D007680:Kidney Neoplasms / Q000235:genetics* / Q000473:pathology; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping; D014407:Tumor Cells, Cultured; D025721:WT1 Proteins; D009396:Wilms Tumor / Q000235:genetics* / Q000473:pathology; D016335:Zinc Fingers / Q000235:genetics*
D004273:DNA, Neoplasm; D004268:DNA-Binding Proteins; D009838:Oligodeoxyribonucleotides; D025721:WT1 Proteins; D008715:Methionine