Abstract
Using purified components in affinity chromatography and blot binding assays, we have found that rat liver lamins A, B, and C can associate in homotypic and heterotypic fashions. Heterotypic A-B and C-B complexes are unusually stable and involve the common amino-terminal domain of lamins A and C, but not their helical "rod" domain. A synthetic peptide, comprising the first 32 amino acid residues of lamins A and C, is able to fully compete with the intact molecules for binding to lamin B. Conversely, heterotypic A-C associations and homotypic A-A and C-C interactions appear significantly weaker than A/C-B binding and do not involve the lamin A and C amino-terminal domain. Homotypic B-B complexes are not formed to any considerable extent unless isolated lamin B subunits are "superphosphorylated" in vitro with protein kinase A. However, when lamins A and C are similarly modified, no changes in their binding specificity can be detected. These data suggest that the nuclear lamina, unlike other multicomponent intermediate filaments, constitutes a nonobligatory heteropolymer. They also indicate that cAMP-dependent phosphorylation of interphase lamin B could cause remodeling of the lamina and establishment of homopolymeric domains.
MeSH Terms
Adenosine Triphosphate/metabolism
Animals
Cell Nucleus/metabolism
Iodine Radioisotopes
Lamin Type A
Lamin Type B
Lamins
Liver/metabolism
Macromolecular Substances
Nuclear Proteins/isolation & purification,metabolism
Phosphorus Radioisotopes
Phosphorylation
Rats
Chemicals
Iodine Radioisotopes
Lamin Type A
Lamin Type B
Lamins
Macromolecular Substances
Nuclear Proteins
Phosphorus Radioisotopes
Adenosine Triphosphate
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Georgatos S D
Laboratory of Cell Biology, Howard Hughes Medical Institute, Rockefeller University, New York, NY 10021.
Stournaras C
Blobel G
References (20)
20 references, click to expand
-
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
Nature. 1970 Aug 15;227(5259):680-5
PMID: 5432063
-
Protein measurement with the Folin phenol reagent.
J Biol Chem. 1951 Nov;193(1):265-75
PMID: 14907713
-
Phosphorylation of intermediate filament proteins by cAMP-dependent protein kinases.
Cell. 1981 Jan;23(1):135-43
PMID: 6260370
-
The nuclear envelope and the architecture of the nuclear periphery.
J Cell Biol. 1981 Dec;91(3 Pt 2):39s-50s
PMID: 7033243
-
Cell type-specific differences in protein composition of nuclear pore complex-lamina structures in oocytes and erythrocytes of Xenopus laevis.
J Mol Biol. 1981 Sep 5;151(1):121-41
PMID: 7328650
-
Nuclear lamina and the structural organization of the nuclear envelope.
Cold Spring Harb Symp Quant Biol. 1982;46 Pt 2:967-78
PMID: 7049540
-
Protein complexes of intermediate-sized filaments: melting of cytokeratin complexes in urea reveals different polypeptide separation characteristics.
Proc Natl Acad Sci U S A. 1983 Dec;80(23):7113-7
PMID: 6196784
-
Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis.
Cell. 1985 May;41(1):177-90
PMID: 3888407
-
Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins.
Nature. 1986 Feb 6-12;319(6053):463-8
PMID: 3453101
-
A cell free system to study reassembly of the nuclear envelope at the end of mitosis.
Cell. 1986 Feb 28;44(4):639-52
PMID: 3948244
-
cDNA sequencing of nuclear lamins A and C reveals primary and secondary structural homology to intermediate filament proteins.
Proc Natl Acad Sci U S A. 1986 Sep;83(17):6450-4
PMID: 3462705
-
The nuclear lamina is a meshwork of intermediate-type filaments.
Nature. 1986 Oct 9-15;323(6088):560-4
PMID: 3762708
-
Nuclear reconstitution in vitro: stages of assembly around protein-free DNA.
Cell. 1987 Jan 30;48(2):205-17
PMID: 3026635
-
Lamin B constitutes an intermediate filament attachment site at the nuclear envelope.
J Cell Biol. 1987 Jul;105(1):117-25
PMID: 3301863
-
Site-specific phosphorylation induces disassembly of vimentin filaments in vitro.
Nature. 1987 Aug 13-19;328(6131):649-52
PMID: 3039376
-
Binding of two desmin derivatives to the plasma membrane and the nuclear envelope of avian erythrocytes: evidence for a conserved site-specificity in intermediate filament-membrane interactions.
Proc Natl Acad Sci U S A. 1987 Oct;84(19):6780-4
PMID: 3477809
-
Teratocarcinoma stem cells and early mouse embryos contain only a single major lamin polypeptide closely resembling lamin B.
Cell. 1987 Nov 6;51(3):383-92
PMID: 3311384
-
Nuclear lamin LI of Xenopus laevis: cDNA cloning, amino acid sequence and binding specificity of a member of the lamin B subfamily.
EMBO J. 1987 Dec 1;6(12):3801-8
PMID: 3428276
-
Phosphorylation of desmin in vitro inhibits formation of intermediate filaments; identification of three kinase A sites in the aminoterminal head domain.
EMBO J. 1988 Jan;7(1):15-20
PMID: 3359992
-
Isolation of nuclear pore complexes in association with a lamina.
Proc Natl Acad Sci U S A. 1975 Mar;72(3):1007-11
PMID: 1055359