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J Biol Chem, Vol. 274, Issue 51, 36520-36526, December 17, 1999
From the MRC Human Genetics Unit, Western General Hospital, Crewe
Road, Edinburgh EH4 2XU, Scotland, United Kingdom
The tumor suppressor gene WT1 encodes
a zinc finger protein, which consists of four C-terminal
C2-H2 zinc fingers of the Krüppel type,
and at the N terminus a Q/P-rich trans-regulatory domain, both
characteristic of transcription factors. However, recent findings
suggest that WT1 may also be involved in a post-transcriptional process. Specifically, WT1 isoforms containing the alternatively spliced exon 9 (+lysine-threonine-serine (KTS)) preferentially associate with nuclear speckles and co-immunoprecipitate splicing antigens (Larsson, S. H., Charlieu, J.-P., Miyagawa, K.,
Engelkamp, D., Rassoulzadegan, M., Ross, A., Cuzin, F., van Heyningen,
V., and Hastie, N. D. (1995) Cell 81, 391-401);
furthermore, WT1 has been shown to interact with the ubiquitous
splicing factor U2AF65 (Davies, R. C., Calvo, C., Larsson, S. H., Lamond, A. I., and Hastie, N. D. (1998) Genes
Dev. 12, 3217-3225) and binds to RNA in vitro
(Caricasole, A., Duarte, A., Larsson, S. H., Hastie, N. D.,
Little, M., Holmes, G., Todorov, I., and Ward, A. (1996) Proc.
Natl. Acad. Sci. U. S. A. 93, 7562-7566; Bardeesy, N., and Pelletier, J. (1998) Nucleic Acids Res. 26, 1784-1792). To
extend these findings, we have fractionated nuclear extracts to see if particles containing WT1 have the properties of ribonucleoprotein (RNP). In summary, WT1 is enriched by oligo(dT) chromatography, as are
U2AF65, the U5 small nuclear RNP-associated protein p116 and hnRNP A1.
Gel filtration and sedimentation profiles suggest that WT1 is present
in RNase-sensitive particles, >2 MDa in size, peaking at ~60 S, and
~1.27 g/cm3 on Nycodenz. Similar results were obtained
from two cell lines expressing WT1, fetal kidneys (day E17), and
transiently transfected cells, suggesting that the presence of WT1
protein in nuclear poly(A)+ RNP is a general aspect of WT1 function.
Presence of WT1, the Wilm's Tumor Suppressor Gene Product, in
Nuclear Poly(A)+ Ribonucleoprotein*
*
This work was supported by a grant from the National Kidney
Research Foundation and the Medical Research Council.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed. Tel.: 0131-3322471;
Fax: 0131-3432620; E-mail: Nick.Hastie@hgu.mrc.ac.uk.
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