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 Lee Wilson, Glenn E. Morris

Research output: Contribution to journalArticle

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

Fingerprint

Emery-Dreifuss Muscular Dystrophy
Lamin Type A
Nuclear Envelope
Nuclear Proteins
Transcription Factors
Lamins
emerin
In Vitro Techniques
RNA Splicing Factors
Site selection
Two-Hybrid System Techniques
Mutation
Proteasome Endopeptidase Complex
Gene Library
Gene expression
Germ Cells
Alanine
Yeast
Muscle
Assays

Keywords

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

ASJC Scopus subject areas

  • Biochemistry

Cite this

Wilkinson, F. L., Holaska, J. M., Zhang, Z., Sharma, A., Manilal, S., Holt, I., ... Morris, G. E. (2003). Emerin interacts in vitro with the splicing-associated factor, YT521-B. European Journal of Biochemistry, 270(11), 2459-2466. https://doi.org/10.1046/j.1432-1033.2003.03617.x

Emerin interacts in vitro with the splicing-associated factor, YT521-B. / Wilkinson, Fiona L.; Holaska, James M.; Zhang, Zhayi; Sharma, Aarti; Manilal, Sushila; Holt, Ian; Stamm, Stefan; Wilson, Katherine Lee; Morris, Glenn E.

In: European Journal of Biochemistry, Vol. 270, No. 11, 06.2003, p. 2459-2466.

Research output: Contribution to journalArticle

Wilkinson, FL, Holaska, JM, Zhang, Z, Sharma, A, Manilal, S, Holt, I, Stamm, S, Wilson, KL & Morris, GE 2003, 'Emerin interacts in vitro with the splicing-associated factor, YT521-B', European Journal of Biochemistry, vol. 270, no. 11, pp. 2459-2466. https://doi.org/10.1046/j.1432-1033.2003.03617.x
Wilkinson FL, Holaska JM, Zhang Z, Sharma A, Manilal S, Holt I et al. Emerin interacts in vitro with the splicing-associated factor, YT521-B. European Journal of Biochemistry. 2003 Jun;270(11):2459-2466. https://doi.org/10.1046/j.1432-1033.2003.03617.x
Wilkinson, Fiona L. ; Holaska, James M. ; Zhang, Zhayi ; Sharma, Aarti ; Manilal, Sushila ; Holt, Ian ; Stamm, Stefan ; Wilson, Katherine Lee ; Morris, Glenn E. / Emerin interacts in vitro with the splicing-associated factor, YT521-B. In: European Journal of Biochemistry. 2003 ; Vol. 270, No. 11. pp. 2459-2466.
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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.",
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AU - Holt, Ian

AU - Stamm, Stefan

AU - Wilson, Katherine Lee

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