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PMID: 9707448 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Detergent-salt resistance of LAP2alpha in interphase nuclei and phosphorylation-dependent association with chromosomes early in nuclear assembly implies functions in nuclear structure dynamics.

The EMBO journal ·Vol. 17 ·No. 16 ·1998-08-17 ·Pages 4887-902

Dechat T, Gotzmann J, Stockinger A, Harris CA, Talle MA, Siekierka JJ, Foisner R

Abstract

Lamina-associated polypeptide (LAP) 2 of the inner nuclear membrane (now LAP2beta) and LAP2alpha are related proteins produced by alternative splicing, and contain a common 187 amino acid N-terminal domain. We show here that, unlike LAP2beta, LAP2alpha behaved like a nuclear non-membrane protein in subcellular fractionation studies and was localized throughout the nuclear interior in interphase cells. It co-fractionated with LAP2beta in nuclear lamina/matrix-enriched fractions upon extraction of nuclei with detergent, salt and nucleases. During metaphase LAP2alpha dissociated from chromosomes and became concentrated around the spindle poles. Furthermore, LAP2alpha was mitotically phosphorylated, and phosphorylation correlated with increased LAP2alpha solubility upon extraction of cells in physiological buffers. LAP2alpha relocated to distinct sites around chromosomes at early stages of nuclear reassembly and intermediarily co-localized with peripheral lamin B and intranuclear lamin A structures at telophase. During in vitro nuclear assembly LAP2alpha was dephosphorylated and assembled into insoluble chromatin-associated structures, and recombinant LAP2alpha was found to interact with chromosomes in vitro. Some LAP2alpha may also associate with membranes prior to chromatin attachment. Altogether the data suggest a role of LAP2alpha in post-mitotic nuclear assembly and in the dynamic structural organization of the nucleus.

MeSH Terms
Animals CHO Cells Cell Nucleus/metabolism,ultrastructure Cricetinae DNA-Binding Proteins Detergents HeLa Cells Humans Interphase Membrane Proteins/metabolism Microscopy, Immunoelectron Mitosis Nuclear Proteins/metabolism Phosphorylation Salts
Chemicals
DNA-Binding Proteins Detergents Membrane Proteins Nuclear Proteins Salts lamina-associated polypeptide 2
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Dechat T
nstitute of Biochemistry and Molecular Cell Biology, Biocenter and Institute of Tumor Biology-Cancer Research, University of Vienna, A-1030 Vienna, Austria.
Gotzmann J
Stockinger A
Harris C A
Talle M A
Siekierka J J
Foisner R
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/17.16.4887
Published
1998-08-17
Pages
4887-902
Language
English
Region
England
NLM ID
8208664
PMCID
PMC1170818
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/17.16.4887 PMC Full Text View journal details
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