title: Assembly of A- and B-type lamins studied in vivo with the baculovirus system.
abstract: We have expressed an A-type lamin (Xenopus lamin A), a probable A-type lamin (Drosophila lamin C), two B-type lamins (Xenopus lamin LI, Drosophila lamin Dmo), and two mutants of Xenopus lamin A in Sf9 cells. All proteins were synthesized at high levels resulting in formation of paracrystals with an axial repeat of 18.5-20.0 nm by A-type lamins; in contrast B-type lamins assembled into aggregates with a fibrillar ultrastructure. Of the four wild-type proteins analyzed only lamin Dmo was found in the nuclear compartment of Sf9 cells in association with the lamina whereas the three other lamins assembled into polymers localized in the cytoplasm as well as the nucleoplasm. The Xenopus lamin A mutant lacking the complete carboxy-terminal tail assembled in the cytoplasm into long filament bundles consisting of fibrils of less than 6 nm diameter. In vitro the non-helical amino-terminal head domain of lamins is required for the formation of 'head-to-tail' polymers. A lamin A mutant lacking this domain could be efficiently extracted from Sf9 cells with physiological buffers containing Triton X-100, demonstrating the importance of this domain for lamin assembly in vivo.
authors.affiliation: Division of Electron Microscopy, Biocenter of the University of Würzburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Klapper
authors.forename: K
authors.initials: K
authors.lastname: Exner
authors.forename: A
authors.initials: A
authors.lastname: Kempf
authors.forename: C
authors.initials: C
authors.lastname: Gehrig
authors.forename: N
authors.initials: N
authors.lastname: Stuurman
authors.forename: P A
authors.initials: PA
authors.lastname: Fisher
authors.forename: G
authors.initials: G
authors.lastname: Krohne
mesh_terms: D000818:Animals; D016367:Baculoviridae; D002460:Cell Line; D004330:Drosophila; D004591:Electrophoresis, Polyacrylamide Gel; D019084:Fluorescent Antibody Technique, Indirect; D005822:Genetic Vectors; D034904:Lamin Type A; D034882:Lamins; D009687:Nuclear Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000648:ultrastructure; D011506:Proteins / Q000032:analysis; D018411:Spodoptera / Q000166:cytology; D014981:Xenopus
chemical_list: D034904:Lamin Type A; D034882:Lamins; D009687:Nuclear Proteins; D011506:Proteins; C440308:lamin C