Published online 30 April 2001. doi:10.1083/jcb.153.3.479
© The Rockefeller University Press,
0021-9525/2001//479 $5.00
The Journal of Cell Biology, Volume 153, Number 3,
, 2001 479-490
Involvement of the Lamin Rod Domain in Heterotypic Lamin Interactions Important for Nuclear Organization
Eric C. Schirmera,
Tinglu Guana, and
Larry Geracea
a Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037
10550 N. Torrey Pines Rd., IMM10, R209, La Jolla, CA 92037.(858) 784-9132(858) 784-8514
lgerace{at}scripps.edu
The nuclear lamina is a meshwork of intermediate-type filament proteins (lamins) that lines the inner nuclear membrane. The lamina is proposed to be an important determinant of nuclear structure, but there has been little direct testing of this idea. To investigate lamina functions, we have characterized a novel lamin B1 mutant lacking the middle
4/5 of its
-helical rod domain. Though retaining only 10 heptads of the rod, this mutant assembles into intermediate filament-like structures in vitro. When expressed in cultured cells, it concentrates in patches at the nuclear envelope. Concurrently, endogenous lamins shift from a uniform to a patchy distribution and lose their complete colocalization, and nuclei become highly lobulated. In vitro binding studies suggest that the internal rod region is important for heterotypic associations of lamin B1, which in turn are required for proper organization of the lamina. Accompanying the changes in lamina structure induced by expression of the mutant, nuclear pore complexes and integral membrane proteins of the inner membrane cluster, principally at the patches of endogenous lamins. Considered together, these data indicate that lamins play a major role in organizing other proteins in the nuclear envelope and in determining nuclear shape.
Key Words: intermediate filament nuclear lamina nuclear pore complex lamina-associated polypeptide nuclear shape
© 2001 The Rockefeller University Press
Abbreviations used in this paper: FTIR, Fourier transform infrared spectroscopy; GFP, green fluorescent protein; HA, hemagglutinin; IF, intermediate filament; INM, inner nuclear membrane; LAP, lamina-associated polypeptide; NE, nuclear envelope; NLS, nuclear localization signal; NPC, nuclear pore complex; NRK, normal rat kidney; WT, wild type.

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