Identification of nuclear localization signals within the zinc fingers of the WT1 tumor suppressor gene product

Bruening, Wendy; Moffett, Peter; Chia, Shea; Heinrich, Günther; Pelletier, Jerry

doi:10.1016/0014-5793(96)00853-8

Cited by 51 publications

(56 citation statements)

References 35 publications

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“…We have experimentally addressed this issue by demonstrating that the NTD-EWS harbors a separate nuclear localization signal (Figure 4). As such EWS/WT1 fusion products contain at least two nuclear localization signals, one with the NTD-EWS and one within zinc ®ngers II-III (Bruening et al, 1996). These results formally demonstrate that EWS/WT1(+KTS) isoforms are nulcear proteins.…”

Section: Discussionsupporting

confidence: 56%

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The DNA binding domains of the WT1 tumor suppressor gene product and chimeric EWS/WT1 oncoprotein are functionally distinct

1998

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The t(11; 22)(p13; q12) translocation associated with desmosplastic small round cell tumor results in a chimeric molecule fusing the amino terminal domain (NTD) of the EWS1 gene to three of the four carboxyterminal zinc ®ngers of the WT1 tumor suppressor gene. Since the DNA binding domains of WT1 and EWS/WT1 are structurally di erent, we have assessed the functional consequences of the EWS/WT1 fusion. We ®nd that the EWS/WT1 protein has a higher binding a nity for a given recognition target than the WT1 product. This is unlike other fusion products involving translocation of the NTD of EWS to DNA binding domains in which DNA binding speci®city and a nity is not changed. We demonstrate that EWS/WT1 is a nuclear protein and that the NTD of EWS contains (a) nuclear localization signal(s). We also ®nd that the integrity of a domain within the WT1 zinc ®ngers, responsible for mediating interaction between WT1 and the transcriptional repressor par-4, is disrupted in the EWS/WT1 fusion product. Deletion analysis of the NTD of EWS indicated that integrity of the entire domain was necessary to achieve full transactivation potential.

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Section: Discussionsupporting

confidence: 56%

“…We have previously shown that WT1 zinc ®ngers II and III harbor a nuclear localization signal (Bruening et al, 1996). Therefore, one consequence of fusing the NTD-EWS to WT1 zinc ®ngers II-IV should be nuclear targeting.…”

Section: The Ntd-ews Harbors a Nuclear Localization Signalmentioning

confidence: 99%

The DNA binding domains of the WT1 tumor suppressor gene product and chimeric EWS/WT1 oncoprotein are functionally distinct

1998

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“…In contrast, expression of WT1(−KTS) clearly inhibited WT1(+KTS) enhancement of CTE function. This might be due to the previously demonstrated ability of the WT1 proteins to form heterodimers through their N-terminal domains (Englert et al 1995;Moffett et al 1995;Reddy et al 1995;Bruening et al 1996). In support of this hypothesis, preliminary experiments suggest that while N-terminal deletion mutants of WT1(+KTS) can still enhance CTE function, expression is no longer inhibited by the WT1(−KTS) protein (Y.-c. Bor, D. Rekosh, and M.L.…”

Section: Discussionmentioning

confidence: 89%

“…This indicates that expression of the WT1(−KTS) protein shows a trans dominant negative effect over the enhancement observed with the +KTS isoform. This observation may relate to the documented ability of WT1 to dimerize via the N terminus (Englert et al 1995;Moffett et al 1995;Reddy et al 1995;Bruening et al 1996).…”

Section: The Wt1(−kts) Protein Inhibits Wt1(+kts) Function In a Dose-mentioning

confidence: 98%

The Wilms’ tumor 1 (WT1) gene (+KTS isoform) functions with a CTE to enhance translation from an unspliced RNA with a retained intron

Bor¹,

Swartz²,

Morrison³

et al. 2006

Genes Dev.

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The Wilms' tumor 1 (WT1) gene plays an important role in mammalian urogenital development, and dysregulation of this gene is observed in many human cancers. Alternative splicing of WT1 RNA leads to the expression of two major protein isoforms, WT1(+KTS) and WT1(−KTS). Whereas WT1(−KTS) acts as a transcriptional regulator, no clear function has been ascribed to WT1(+KTS), despite the fact that this protein is crucial for normal development. Here we show that WT1(+KTS) functions to enhance expression from RNA possessing a retained intron and containing either a cellular or viral constitutive transport element (CTE). WT1(+KTS) expression increases the levels of unspliced RNA containing a CTE and specifically promotes the association of this RNA with polyribosomes. These studies provide further support for links between different steps in RNA metabolism and for the existence of post-transcriptional operons.

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“…It is responsible for transcription and RNA processing (22). However, WT1 can be detected in the cytoplasm of various cell lines including breast cancer and shuttles the nucleus and the cytoplasm (23,24).…”

Section: -Dementioning

confidence: 99%

Proteomics analysis of siRNA-mediated silencing of Wilms’ tumor 1 in the MDA-MB-468 breast cancer cell line

et al. 2014

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Abstract. The Wilms' tumor 1 (WT1) gene encodes a zinc finger which appears to be a transcriptional activator or repressor for many genes involved in cell differentiation, growth and apoptosis. In order to determine the relationship between WT1 and related proteins, WT1 was silenced with small interfering RNA (siRNA) and the protein expression pattern was analyzed by proteomics analysis including one-dimensional gel electrophoresis (1-DE) and LC-MS/MS mass spectrometry. The results revealed that 14 proteins were expressed in WT1-silenced cells (siRNA WT1 ) and 12 proteins were expressed in the WT1-expressing cells (siRNA neg ), respectively. These proteins may be classified by their functions in apoptosis, cell signaling, protein folding, gene expression, redox-regulation, transport, structural and unknown functions. Mitogaligin, an apoptosis-related molecule, was identified when WT1 was silenced while the proteins related to the signaling pathway were detected in both siRNA neg and siRNA WT1 but the type of proteins were different. For example, the IBtK protein and the SH2 domain-containing protein were present in siRNA WT1 conditions, while the platelet-derived growth factor receptor α (PDGFRA) and Rho guanine nucleotide exchange factor 1 (Rho-GEF 1) were expressed in siRNA neg . Of these, Rho-GEF was selected for validation by western blot analysis and demonstrated to be present only in the presence of WT1. In conclusion, WT1 is related to mitogaligin via EGFR and behaves as an anti-apoptotic molecule. Moreover, WT1 may be associated with PDGFRA and Rho-GEF 1 that activates proliferation in MDA-MB-468 cells.

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Identification of nuclear localization signals within the zinc fingers of the WT1 tumor suppressor gene product

Cited by 51 publications

References 35 publications

The DNA binding domains of the WT1 tumor suppressor gene product and chimeric EWS/WT1 oncoprotein are functionally distinct

The DNA binding domains of the WT1 tumor suppressor gene product and chimeric EWS/WT1 oncoprotein are functionally distinct

The Wilms’ tumor 1 (WT1) gene (+KTS isoform) functions with a CTE to enhance translation from an unspliced RNA with a retained intron

Proteomics analysis of siRNA-mediated silencing of Wilms’ tumor 1 in the MDA-MB-468 breast cancer cell line

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