title: Interphase phosphorylation of lamin A.
abstract: Nuclear lamins form the major structural elements that comprise the nuclear lamina. Loss of nuclear structural integrity has been implicated as a key factor in the lamin A/C gene mutations that cause laminopathies, whereas the normal regulation of lamin A assembly and organization in interphase cells is still undefined. We assumed phosphorylation to be a major determinant, identifying 20 prime interphase phosphorylation sites, of which eight were high-turnover sites. We examined the roles of these latter sites by site-directed mutagenesis, followed by detailed microscopic analysis - including fluorescence recovery after photobleaching, fluorescence correlation spectroscopy and nuclear extraction techniques. The results reveal three phosphorylation regions, each with dominant sites, together controlling lamin A structure and dynamics. Interestingly, two of these interphase sites are hyper-phosphorylated in mitotic cells and one of these sites is within the sequence that is missing in progerin of the Hutchinson-Gilford progeria syndrome. We present a model where different phosphorylation combinations yield markedly different effects on the assembly, subunit turnover and the mobility of lamin A between, and within, the lamina, the nucleoplasm and the cytoplasm of interphase cells.
authors.affiliation: Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, FIN-20521 Turku, Finland Department of Biosciences, Åbo Akademi University, FIN-20520 Turku, Finland Department of Pathology, Sapporo Medical University, Sapporo, Hokkaido 060-8556, Japan.
authors.forename: Vitaly
authors.initials: V
authors.lastname: Kochin
authors.affiliation: Northwestern University Feinberg School of Medicine, Department of Cell and Molecular Biology, Chicago, IL 60611, USA.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Shimi
authors.affiliation: Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, FIN-20521 Turku, Finland Department of Biosciences, Åbo Akademi University, FIN-20520 Turku, Finland.
authors.forename: Elin
authors.initials: E
authors.lastname: Torvaldson
authors.affiliation: Northwestern University Feinberg School of Medicine, Department of Cell and Molecular Biology, Chicago, IL 60611, USA.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Adam
authors.affiliation: Northwestern University Feinberg School of Medicine, Department of Cell and Molecular Biology, Chicago, IL 60611, USA.
authors.forename: Anne
authors.initials: A
authors.lastname: Goldman
authors.affiliation: Cellular Informatics Laboratory, RIKEN, Wako-shi, Saitama 351-0198, Japan.
authors.forename: Chan-Gi
authors.initials: CG
authors.lastname: Pack
authors.affiliation: Northwestern University Feinberg School of Medicine, Department of Cell and Molecular Biology, Chicago, IL 60611, USA.
authors.forename: Johanna
authors.initials: J
authors.lastname: Melo-Cardenas
authors.affiliation: Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, FIN-20521 Turku, Finland.
authors.forename: Susumu Y
authors.initials: SY
authors.lastname: Imanishi
authors.affiliation: Northwestern University Feinberg School of Medicine, Department of Cell and Molecular Biology, Chicago, IL 60611, USA.
authors.forename: Robert D
authors.initials: RD
authors.lastname: Goldman
authors.affiliation: Turku Centre for Biotechnology, University of Turku and Åbo Akademi University, FIN-20521 Turku, Finland Department of Biosciences, Åbo Akademi University, FIN-20520 Turku, Finland john.eriksson@abo.fi.
authors.forename: John E
authors.initials: JE
authors.lastname: Eriksson
mesh_terms: D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D006367:HeLa Cells; D006801:Humans; D007399:Interphase; D034904:Lamin Type A / Q000378:metabolism*; D008969:Molecular Sequence Data; D034881:Nuclear Lamina / Q000378:metabolism; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases; D021381:Protein Transport; D014157:Transcription Factors; D000092468:Bromodomain Containing Proteins
chemical_list: D034904:Lamin Type A; D017346:Protein Serine-Threonine Kinases; D014157:Transcription Factors; C098415:BRD2 protein, human; D000092468:Bromodomain Containing Proteins; C494847:LMNA protein, human
keywords: Intermediate filament; Lamin A; Phosphorylation; Sequestration; Signaling
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