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

Oncogene. 2008 Jan 24;27(5):653-62. doi: 10.1038/sj.onc.1210688. Epub 2007 Jul 30.

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-alpha (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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • COS Cells
  • Chlorocebus aethiops
  • Gene Expression Regulation
  • Haplorhini
  • HeLa Cells
  • Homeodomain Proteins / metabolism*
  • Humans
  • LIM-Homeodomain Proteins
  • Neoplasm Proteins / metabolism*
  • Promoter Regions, Genetic
  • Protein Interaction Domains and Motifs
  • Proto-Oncogene Proteins / metabolism
  • Repressor Proteins / metabolism*
  • Sarcoma, Synovial / metabolism*
  • Transcription Factors / metabolism*
  • Transcription, Genetic

Substances

  • Homeodomain Proteins
  • LHX4 protein, human
  • LIM-Homeodomain Proteins
  • Neoplasm Proteins
  • Proto-Oncogene Proteins
  • Repressor Proteins
  • SS18 protein, human
  • Transcription Factors
  • synovial sarcoma X breakpoint proteins