Immunoelectron microscopic studies on centromere-kinetochore complexes detached from chromosomes |
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Authors: | N. Paweletz E. -M. Finze D. Schroeter M. Enulescu M. Knehr |
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Affiliation: | (1) the Research Program IV, Division 0430, German Cancer Research Center, Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany |
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Abstract: | The centromere-kinetochore complexes of Chinese hamster ovary (CHO) cells were detached and separated from the condensed chromatin by treatment with hydroxyurea and caffeine. By labelling the complex for immunoelectron microscopy (immuno-EM) with a mixture of antibodies against centromere proteins (anti-CENP-A,-B, -C) in some cells, we could demonstrate complete detachment of the complexes. No remnants were left at the bulk of condensed chromatin in these cells. In some mitotic cells complex and chromatin were found side by side. It could be shown that the fine structure of the separated material of the complex differs significantly from that of the rest of chromatin. The complex consists of proteins and DNA. This leads us to suppose that the organization of chromatin in the centromere-kinetochore complex is different. |
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Keywords: | caffeine centromere-kinetochore complex CHO cells chromatin hydroxyurea immunoelectron microscopy |
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