title: Identification of differential protein interactors of lamin A and progerin.
abstract: The nuclear lamina is an interconnected meshwork of intermediate filament proteins underlying the nuclear envelope. The lamina is an important regulator of nuclear structural integrity as well as nuclear processes, including transcription, DNA replication and chromatin remodeling. The major components of the lamina are A- and B-type lamins. Mutations in lamins impair lamina functions and cause a set of highly tissue-specific diseases collectively referred to as laminopathies. The phenotypic diversity amongst laminopathies is hypothesized to be caused by mutations affecting specific protein interactions, possibly in a tissue-specific manner. Current technologies to identify interaction partners of lamin A and its mutants are hampered by the insoluble nature of lamina components. To overcome the limitations of current technologies, we developed and applied a novel, unbiased approach to identify lamin A-interacting proteins. This approach involves expression of the high-affinity OneSTrEP-tag, precipitation of lamin-protein complexes after reversible protein cross-linking and subsequent protein identification by mass spectrometry. We used this approach to identify in mouse embryonic fibroblasts and cardiac myocyte NklTAg cell lines proteins that interact with lamin A and its mutant isoform progerin, which causes the premature aging disorder Hutchinson-Gilford progeria syndrome (HGPS). We identified a total of 313 lamina-interacting proteins, including several novel lamin A interactors, and we characterize a set of 35 proteins which preferentially interact with lamin A or progerin.
authors.affiliation: Center for Heart Failure Research, Cardiovascular Research Institute Maastricht, The Netherlands.
authors.forename: Nard
authors.initials: N
authors.lastname: Kubben
authors.forename: Jan Willem
authors.initials: JW
authors.lastname: Voncken
authors.forename: Jeroen
authors.initials: J
authors.lastname: Demmers
authors.forename: Chantal
authors.initials: C
authors.lastname: Calis
authors.forename: Geert
authors.initials: G
authors.lastname: van Almen
authors.forename: Yigal
authors.initials: Y
authors.lastname: Pinto
authors.forename: Tom
authors.initials: T
authors.lastname: Misteli
mesh_terms: D000818:Animals; D002460:Cell Line; D005347:Fibroblasts / Q000378:metabolism; D055786:Gene Knockout Techniques; D006801:Humans; D034904:Lamin Type A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry; D051379:Mice; D009154:Mutation; D032383:Myocytes, Cardiac / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011371:Progeria / Q000378:metabolism / Q000473:pathology; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D034904:Lamin Type A; D009687:Nuclear Proteins; D020033:Protein Isoforms; D011498:Protein Precursors; C090764:prelamin A