Unassembled (soluble) vimentin in human myeloid leukemia cell line HL60.
The intermediate filament proteins which include vimentin, desmin, and the keratins are one of three major classes of cytoskeletal proteins in eukaryotic cells. In this study we found that most of the vimentin of undifferentiated HL60 and cells induced to differentiate either along the monocytoid pathway by 12-O-tetradecanoylphorbol-13-acetate (TPA) or along the granulocytic pathway by retinoic acid was soluble in a buffer containing 1% Triton X-100/0.6 mol/l KCl in which the intermediate filament proteins usually are not soluble. HL60 vimentin separated on polyacrylamide gel electrophoresis into two proteins of Mr 55,000 and 54,000 that we detected by immunoblotting. The Mr 55,000 species was the major form in undifferentiated HL60 cells and cells induced by retinoic acid. The distribution of both forms of vimentin changed during induction of differentiation by TPA and after 24 h the Mr 54,000 species was predominant. After an additional 24 h exposure to TPA the relative levels of the two forms of vimentin approached equivalence and a high level of vimentin degradation products was seen. These results suggest that TPA may increase vimentin degradation along a pathway that has a Mr 54,000 intermediate. In addition, the high levels of soluble vimentin in HL60 cells suggests that these cells may be a good model for studying components involved in vimentin assembly.
Takahashi
N
N
Laboratory of Biological Chemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892.
Breitman
T R
TR
D000911:Antibodies, Monoclonal; D002454:Cell Differentiation / Q000187:drug effects; D003893:Desmin / Q000032:analysis / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007633:Keratins / Q000032:analysis / Q000378:metabolism; D007951:Leukemia, Myeloid / Q000378:metabolism / Q000473:pathology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014212:Tretinoin / Q000494:pharmacology; D014407:Tumor Cells, Cultured / Q000737:chemistry; D014746:Vimentin / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism
D000911:Antibodies, Monoclonal; D003893:Desmin; D014746:Vimentin; D014212:Tretinoin; D007633:Keratins; D013755:Tetradecanoylphorbol Acetate