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Presence of WT1, the Wilm's tumor suppressor gene product, in nuclear poly(A)+ ribonucleoprotein

Ladomery, Michael R.; Hastie, Nicholas D.; Ladomery, Michael; Slight, Joan; Hastie, Nicholas; Mc Ghee, Sharon

Authors

Michael R. Ladomery

Nicholas D. Hastie

Joan Slight

Nicholas Hastie

Sharon Mc Ghee



Abstract

The tumor suppressor gene WT1 encodes a zinc finger protein, which consists of four C-terminal C2-H2 zinc fingers of the Kruppel 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 coimmunoprecipitate 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.

Citation

Hastie, N. D., Ladomery, M. R., Ladomery, M., Slight, J., Mc Ghee, S., & Hastie, N. (1999). Presence of WT1, the Wilm's tumor suppressor gene product, in nuclear poly(A)+ ribonucleoprotein. Journal of Biological Chemistry, 274(51), 36520-36526. https://doi.org/10.1074/jbc.274.51.36520

Journal Article Type Article
Publication Date Dec 17, 1999
Journal Journal of Biological Chemistry
Print ISSN 0021-9258
Publisher American Society for Biochemistry and Molecular Biology
Peer Reviewed Peer Reviewed
Volume 274
Issue 51
Pages 36520-36526
DOI https://doi.org/10.1074/jbc.274.51.36520
Public URL https://uwe-repository.worktribe.com/output/1096171
Publisher URL http://dx.doi.org/10.1074/jbc.274.51.36520