The prometaphase configuration and chromosome order in early mitosis

Abstract

We have examined early mitotic stages in HeLa cells, mouse 3T3 fibroblasts and mitogen-activated mouse lymphocytes by immunofluorescence labelling with anti-tubulin and anti-centromere. Chromatin organization was monitored with the DNA-specific fluorochrome Hoechst 33258. This approach has led us to identify a modified Rabl array of chromosomes and spindle microtubules early in mitosis that is distinct from that at metaphase, and which we have called ‘the prometaphase configuration’. In the configuration, chromosomes are oriented so that telomeres are clustered at the outer surface, whereas centromeres are clustered inside the configuration, at the surface of a hollow spindle. Observations on cells earlier in mitosis indicate that the configuration is presaged by the spatial relationship between chromosomes and cytoplasmic microtubules in prophase and early prometaphase. We propose a model in which the prometaphase configuration represents an important step linking prophase and metaphase, serving to translate interphase spatial and intragenomic order into order at the metaphase plate.