Immunoaffinity purification and functional characterization of interphase and meiotic Drosophila nuclear lamin isoforms.
Three isoforms of a single nuclear lamin have been identified in Drosophila. Two, lamins Dm1 and Dm2, are present during interphase and are apparently in equilibrium with each other in vivo. The third, lamin Dmmit, is found in cells that have undergone nuclear envelope breakdown, either during meiosis or mitosis. All three isoforms were purified under nondenaturing conditions using a novel technique of immunoaffinity chromatography and their in vitro activities were examined. Interphase lamins Dm1 and Dm2 can assemble into filaments at physiologic ionic strength; assembly is reversible upon addition of concentrated NaCl. Negative staining of filaments formed in vitro shows long, unbranched bundles approximately 20 nm in diameter. Addition of specific antilamin antibodies blocks in vitro assembly completely. In contrast with lamins Dm1 and Dm2, lamin Dmmit remains soluble at physiologic ionic strength. These observations are consistent with the notion that lamina disassembly in vivo is due, at least in part, to changes in properties of the lamins themselves.
Lin
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Department of Pharmacological Sciences, State University of New York, Stony Brook, 11794-8651.
Fisher
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PA
D000818:Animals; D002846:Chromatography, Affinity; D004331:Drosophila melanogaster; D004591:Electrophoresis, Polyacrylamide Gel; D007158:Immunologic Techniques; D007399:Interphase; D034882:Lamins; D046911:Macromolecular Substances; D008540:Meiosis; D008854:Microscopy, Electron; D009687:Nuclear Proteins / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D009994:Osmolar Concentration; D011485:Protein Binding; D012995:Solubility
D034882:Lamins; D046911:Macromolecular Substances; D009687:Nuclear Proteins