title: Emerin interacts in vitro with the splicing-associated factor, YT521-B.
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 PA28 gamma 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.
authors.affiliation: Biochemistry Group, North-East Wales Institute, Wrexham, UK.
authors.forename: Fiona L
authors.initials: FL
authors.lastname: Wilkinson
authors.forename: James M
authors.initials: JM
authors.lastname: Holaska
authors.forename: Zhayi
authors.initials: Z
authors.lastname: Zhang
authors.forename: Aarti
authors.initials: A
authors.lastname: Sharma
authors.forename: Sushila
authors.initials: S
authors.lastname: Manilal
authors.forename: Ian
authors.initials: I
authors.lastname: Holt
authors.forename: Stefan
authors.initials: S
authors.lastname: Stamm
authors.forename: Katherine L
authors.initials: KL
authors.lastname: Wilson
authors.forename: Glenn E
authors.initials: GE
authors.lastname: Morris
mesh_terms: D001665:Binding Sites; D002467:Cell Nucleus / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D029721:Drosophila Proteins; D015723:Gene Library; D017930:Genes, Reporter; D006801:Humans; D066298:In Vitro Techniques; D036341:Intercellular Signaling Peptides and Proteins; D034904:Lamin Type A / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000378:metabolism; D020389:Muscular Dystrophy, Emery-Dreifuss / Q000378:metabolism; D009206:Myocardium / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism; D016366:Open Reading Frames; D011233:Precipitin Tests; D046988:Proteasome Endopeptidase Complex; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000378:metabolism; D012326:RNA Splicing; D000072260:RNA Splicing Factors; D012333:RNA, Messenger / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D013946:Thymopoietins / Q000737:chemistry / Q000378:metabolism*; D013997:Time Factors; D020798:Two-Hybrid System Techniques
chemical_list: D018076:DNA, Complementary; D029721:Drosophila Proteins; C471550:Gcl protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D034904:Lamin Type A; D008565:Membrane Proteins; D009097:Multienzyme Complexes; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D000072260:RNA Splicing Factors; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D013946:Thymopoietins; C490769:YTHDC1 protein, human; C092710:emerin; C076187:gcl protein, Drosophila; D012313:RNA; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex