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The C terminus of the synovial sarcoma-associated SSX proteins interacts with the LIM homeobox protein LHX4

Oncogene volume 27, pages 653–662 (2008)Cite this article

Abstract

As a result of the synovial sarcoma-associated t(X;18) translocation, the SS18 gene on chromosome 18 is fused to either one of the three closely related SSX genes on the X chromosome. The SS18 protein is thought to act as a transcriptional co-activator, whereas the SSX proteins are thought to act as transcriptional corepressors. The main SSX-repression domain is located in its C terminus, a domain that is retained in the respective SS18–SSX fusion proteins. Both the SS18 and SSX proteins lack DNA-binding domains. Previously, we found that the SS18 and SS18–SSX fusion proteins may be tethered to DNA targets via the SS18-interacting protein AF10. Here, we set out to isolate proteins that interact with the SSX C-terminal repression domain using a yeast two-hybrid interaction trap. Of the positive clones isolated, two corresponded to the LIM homeobox protein LHX4, a DNA-binding protein that is involved in transcription regulation. An endogenous interaction was subsequently established in mammalian cells via colocalization and coimmunoprecipitation of the respective proteins. Interestingly, the LHX4 gene was previously found to be deregulated in various human leukemias. In addition, it was previously found that LIM homeobox proteins may bind to and activate the glycoprotein-α (CGA) promoter. Using LHX4 chromatin immunoprecipitation and CGA-promoter assays, we found that endogenous LHX4 binds to the CGA promoter and that LHX4-mediated CGA activation is enhanced by the SS18–SSX protein, but not by the SSX protein. Taken together, we conclude that this novel protein – protein interaction may have direct consequences for the (de)regulation of SSX and/or SS18–SSX target genes and, thus, for the development of human synovial sarcomas.

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Acknowledgements

We thank Jack Fransen (Department of Cell Biology, Nijmegen Centre for Molecular Life Sciences) for assistance with the confocal laser scanning microscopy. This work was supported by a grant from the Dutch Cancer Society (KWF kankerbestrijding).

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  1. Department of Human Genetics, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands

    D R H de Bruijn, A H A van Dijk, M P Willemse & A Geurts van Kessel

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  1. D R H de Bruijn
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Correspondence to A Geurts van Kessel.

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de Bruijn, D., van Dijk, A., Willemse, M. et al. The C terminus of the synovial sarcoma-associated SSX proteins interacts with the LIM homeobox protein LHX4. Oncogene 27, 653–662 (2008). https://doi.org/10.1038/sj.onc.1210688

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