In Vitro Posttranslational Modification of Lamin B Cloned from a Human T-Cell Line
| Autor: | K M, Pollard, E K, Chan, B J, Grant, K F, Sullivan, E M, Tan, C A, Glass |
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| Rok vydání: | 1990 |
| Předmět: |
animal structures
Base Sequence Lamin Type B integumentary system Molecular Sequence Data Antibodies Monoclonal Nuclear Proteins DNA Cell Biology Lamins Cell Compartmentation Cell Line Blotting Southern Protein Biosynthesis Humans Electrophoresis Gel Two-Dimensional Amino Acid Sequence Cloning Molecular Protein Processing Post-Translational Molecular Biology Research Article |
| Zdroj: | Molecular and Cellular Biology. 10:2164-2175 |
| ISSN: | 1098-5549 |
| Popis: | Autoimmune diseases are characterized by spontaneously occurring autoantibodies which have proven to be useful reagents for the characterization of specific nuclear proteins. Using a monoclonal autoantibody (72B9) derived from a murine lupus strain, we have cloned a cDNA from the human T-cell line MOLT-4, which encodes nuclear lamin B. The identity of the encoded protein as lamin B was established by both biochemical and immunological criteria. Inspection of the deduced amino acid sequence of lamin B revealed the presence in coil 1B of the alpha-helical domain of a leucine heptad repeat region. Analysis of mRNA in HL60 and MOLT-4 cells, which express only lamin B, or HeLa cells, which express all three major lamins (A, B, and C), together with the comigration of in vitro-translated product with isolated HeLa cell lamin B by two-dimensional gel electrophoresis, suggests that a single lamin B is expressed in mammalian somatic cells. In vitro translation with the cDNA clone revealed an EDTA-sensitive posttranslational modification which resulted in an increase in the apparent molecular weight to that equivalent to the native in vivo-synthesized lamin B protein. This in vitro modification included incorporation of a product of mevalonolactone and required an intact carboxy terminus. |
| Databáze: | OpenAIRE |
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