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Impairment of MAD2B-PRCC interaction in mitotic checkpoint defective t(X;1)-positive renal cell carcinomas
M A Weterman1, J J van Groningen, L Tertoolen
1Department of Human Genetics, University Medical Center Nijmegen, 6500 HB Nijmegen, The Netherlands. A.Geurtsvankessel@antrg.azn.nl
Abstract:
The papillary renal cell carcinoma (RCC)-associated (X;1)(p11;q21) translocation fuses the genes PRCC and TFE3 and leads to cancer by an unknown molecular mechanism. We here demonstrate that the mitotic checkpoint protein MAD2B interacts with PRCC. The PRCCTFE3 fusion protein retains the MAD2B interaction domain, but this interaction is impaired. In addition, we show that two t(X;1)-positive RCC tumor cell lines are defective in their mitotic checkpoint. Transfection of PRCCTFE3, but not the reciprocal product TFE3PRCC, disrupts the mitotic checkpoint in human embryonic kidney cells. Our results suggest a dominant-negative effect of the PRCCTFE3 fusion gene leading to a mitotic checkpoint defect as an early event in papillary RCCs.
Insights
The PRCC-TFE3 fusion gene in papillary renal cell carcinoma (RCC) impairs the mitotic checkpoint by disrupting MAD2B interaction. This defect may be an early event in papillary RCC development.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Papillary renal cell carcinoma (RCC) is associated with the (X;1)(p11;q21) translocation, fusing PRCC and TFE3 genes.
- The molecular mechanisms driving tumorigenesis in papillary RCC remain largely unknown.
Purpose of the Study:
- To investigate the molecular mechanism by which the PRCC-TFE3 fusion contributes to papillary RCC development.
- To explore the role of the mitotic checkpoint in PRCC-TFE3-driven tumorigenesis.
Main Methods:
- Co-immunoprecipitation assays to assess protein interactions.
- Mitotic checkpoint function analysis in cancer cell lines and transfected cells.
- Western blotting to detect protein expression and interaction.
Main Results:
- The mitotic checkpoint protein MAD2B interacts with the wild-type PRCC protein.
- The PRCC-TFE3 fusion protein retains the MAD2B interaction domain, but the interaction is impaired.
- PRCC-TFE3 transfection disrupts the mitotic checkpoint in human embryonic kidney cells, suggesting a dominant-negative effect.
Conclusions:
- The PRCC-TFE3 fusion gene likely causes papillary RCC through a dominant-negative disruption of the mitotic checkpoint.
- Impaired MAD2B interaction and subsequent mitotic checkpoint defects are proposed as early events in papillary RCC pathogenesis.