TY - JOUR
T1 - Nuclear transport of wilms′ tumour protein Wt1 involves importins α and β
AU - Depping, Reinhard
AU - Schindler, Susann Gaby
AU - Jacobi, Charlotte
AU - Kirschner, Karin M.
AU - Scholz, Holger
N1 - Copyright:
Copyright 2019 Elsevier B.V., All rights reserved.
PY - 2012
Y1 - 2012
N2 - Background/Aims: Wilms′ tumour protein, Wt1, is a zinc finger molecule, which is required for normal embryonic development. Mutations of the WT1 gene can give rise to childhood cancer of the kidneys. Different Wt1 isoforms exist, which function either as transcription factors or have a presumed role in mRNA processing. Previous studies suggested that Wt1 undergoes nucleocytoplasmic shuttling, and cytoplasmic Wt1 was higher in malignant than in normal cells. The aim of this study was to analyse the molecular pathways along which Wt1 shuttles between the cytoplasm and nucleus. Methods: Interaction of Wt1 protein with various importin α subtypes and importin β was assessed in pull-down assays and co-immunoprecipitation experiments. Nuclear localisation signals (NLS) were identified by combining site-directed mutagenesis with subcellular immunodetection of the transfected Wt1 variants. Results: Wt1(+/-KTS) proteins were found to interact with importin α1 and importin β in vitro and in living cells in vivo. A NLS that was necessary and sufficient for nuclear import could be mapped to the third Wt1 zinc finger. Mutation of this NLS strongly weakened binding of Wt1 to importins. Conclusion: Nuclear translocation of Wilms′ tumour protein involves importins α and β, and a NLS in the third zinc finger.
AB - Background/Aims: Wilms′ tumour protein, Wt1, is a zinc finger molecule, which is required for normal embryonic development. Mutations of the WT1 gene can give rise to childhood cancer of the kidneys. Different Wt1 isoforms exist, which function either as transcription factors or have a presumed role in mRNA processing. Previous studies suggested that Wt1 undergoes nucleocytoplasmic shuttling, and cytoplasmic Wt1 was higher in malignant than in normal cells. The aim of this study was to analyse the molecular pathways along which Wt1 shuttles between the cytoplasm and nucleus. Methods: Interaction of Wt1 protein with various importin α subtypes and importin β was assessed in pull-down assays and co-immunoprecipitation experiments. Nuclear localisation signals (NLS) were identified by combining site-directed mutagenesis with subcellular immunodetection of the transfected Wt1 variants. Results: Wt1(+/-KTS) proteins were found to interact with importin α1 and importin β in vitro and in living cells in vivo. A NLS that was necessary and sufficient for nuclear import could be mapped to the third Wt1 zinc finger. Mutation of this NLS strongly weakened binding of Wt1 to importins. Conclusion: Nuclear translocation of Wilms′ tumour protein involves importins α and β, and a NLS in the third zinc finger.
UR - http://www.scopus.com/inward/record.url?scp=84859848486&partnerID=8YFLogxK
U2 - 10.1159/000337603
DO - 10.1159/000337603
M3 - Journal articles
C2 - 22415091
AN - SCOPUS:84859848486
VL - 29
SP - 223
EP - 232
JO - Cellular Physiology and Biochemistry
JF - Cellular Physiology and Biochemistry
SN - 1015-8987
IS - 1-2
ER -