title: Prelamin A processing, accumulation and distribution in normal cells and laminopathy disorders.
abstract: Lamin A is part of a complex structural meshwork located beneath the nuclear envelope and is involved in both structural support and the regulation of gene expression. Lamin A is initially expressed as prelamin A, which contains an extended carboxyl terminus that undergoes a series of post-translational modifications and subsequent cleavage by the endopeptidase ZMPSTE24 to generate lamin A. To facilitate investigations of the role of this cleavage in normal and disease states, we developed a monoclonal antibody (PL-1C7) that specifically recognizes prelamin A at the intact ZMPSTE24 cleavage site, ensuring prelamin A detection exclusively. Importantly, PL-1C7 can be used to determine prelamin A localization and accumulation in cells where lamin A is highly expressed without the use of exogenous fusion proteins. Our results show that unlike mature lamin A, prelamin A accumulates as discrete and localized foci at the nuclear periphery. Furthermore, whereas treatment with farnesylation inhibitors of cells overexpressing a GFP-prelamin A fusion protein results in the formation of large nucleoplasmic clumps, these aggregates are not observed upon similar treatment of cells expressing endogenous prelamin A or in cells lacking ZMPSTE24 expression and/or activity. Finally, we show that specific laminopathy-associated mutations exhibit both positive and negative effects on prelamin A accumulation, indicating that these mutations affect prelamin A processing efficiency in different manners.
authors.affiliation: a Basic Science Division, Fred Hutchinson Cancer Research Center , Seattle , WA , USA.|c Instituto Fisiología Celular, Universidad Nacional Autónoma de México , Mexico City , Mexico.
authors.forename: Andrea
authors.initials: A
authors.lastname: Casasola
authors.affiliation: a Basic Science Division, Fred Hutchinson Cancer Research Center , Seattle , WA , USA.
authors.forename: David
authors.initials: D
authors.lastname: Scalzo
authors.affiliation: a Basic Science Division, Fred Hutchinson Cancer Research Center , Seattle , WA , USA.
authors.forename: Vivek
authors.initials: V
authors.lastname: Nandakumar
authors.affiliation: a Basic Science Division, Fred Hutchinson Cancer Research Center , Seattle , WA , USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Halow
authors.affiliation: c Instituto Fisiología Celular, Universidad Nacional Autónoma de México , Mexico City , Mexico.
authors.forename: Félix
authors.initials: F
authors.lastname: Recillas-Targa
authors.affiliation: a Basic Science Division, Fred Hutchinson Cancer Research Center , Seattle , WA , USA.|b Department of Radiation Oncology , University Washington School of Medicine , Seattle , WA , USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Groudine
authors.affiliation: a Basic Science Division, Fred Hutchinson Cancer Research Center , Seattle , WA , USA.
authors.forename: Héctor
authors.initials: H
authors.lastname: Rincón-Arano
mesh_terms: D000818:Animals; D006367:HeLa Cells; D006801:Humans; D034904:Lamin Type A / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009154:Mutation; D011371:Progeria / Q000235:genetics / Q000378:metabolism*; D017368:Protein Prenylation; D021381:Protein Transport
chemical_list: D034904:Lamin Type A; D008565:Membrane Proteins; C090764:prelamin A; D008666:Metalloendopeptidases; C491250:Zmpste24 protein, mouse; C120130:ZMPSTE24 protein, human
keywords: ZMPSTE24; intracellular flow cytometry (IFC); lamin A; monoclonal antibody; nuclear envelope; post-translational processing; prelamin A; progeriod syndromes