Emerin interacts in vitro with the splicing-associated factor, YT521-B

Fiona L. Wilkinson, James M. Holaska, Zhayi Zhang, Aarti Sharma, Sushila Manilal, Ian Holt, Stefan Stamm, Katherine L. Wilson, Glenn E. Morris

Research output: Contribution to journalArticlepeer-review

Abstract

Emerin is a nuclear membrane protein that interacts with lamin A/C at the nuclear envelope. Mutations in either emerin or lamin A/C cause Emery-Dreifuss muscular dystrophy (EDMD). The functions of emerin are poorly understood, but EDMD affects mainly skeletal and cardiac muscle. We used a high-stringency yeast two-hybrid method to screen a human heart cDNA library, with full-length emerin as bait. Four out of five candidate interactors identified were nuclear proteins: lamin A, splicing factor YT521-B, proteasome subunit PA28γ and transcription factor vav-1. Specific binding between emerin and the functional C-terminal domain of YT521-B was confirmed by pull-down assays and biomolecular interaction analysis (BIAcore). Inhibition by emerin of YT521-B-dependent splice site selection in vivo suggests that the interaction is physiologically significant. A 'bipartite' binding site for YT521-B in emerin was identified using alanine substitution or disease-associated mutations in emerin. The transcription factor GCL (germ cell-less) has previously been shown to bind to the same site. The results are consistent with an emerging view that lamins and lamina-associated proteins, like emerin, have a regulatory role, as well as a structural role in the nucleus. YT521-B joins a growing list of candidates for a role in a gene expression model of the pathogenesis of EDMD.

Original languageEnglish (US)
Pages (from-to)2459-2466
Number of pages8
JournalEuropean Journal of Biochemistry
Volume270
Issue number11
DOIs
StatePublished - Jun 2003

Keywords

  • Emery-Dreifuss muscular dystrophy
  • Gene regulation
  • Lamin
  • RNA splicing
  • Yeast two-hybrid

ASJC Scopus subject areas

  • Biochemistry

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