The kinetochore is a structurally differentiated site on mitotic chromosomes to which spindle microtubules (MTs) are attached. In mammalian cells, the kinetochore is organized into a trilamellar plate and is morphologically distinct from the centromere. Although kinetochores and centromeres are morphologically and biochemically distinct regions, they are functionally linked and necessary for normal chromosome movement and segregation. Recent biochemical and immunocytochemical studies suggest that the kinetochore is composed of several polypeptides, DNA, and possibly RNA. The kinetochore plates are composed of tubulin and two antigens of 17 Kd and 80 Kd, as detected by scleroderma CREST antiserum. Colcemid, a MT inhibitor, also causes reversible rearrangements of kinetochore structure. Mitomycin C binds to heterochromatin and causes the trilamellar plates to become detached from the chromosome. Diethylstilbestrol (DES), a synthetic estrogen, inhibits mitosis in mammalian cells and causes chromosome lagging or malorientation during recovery. Electron microscopy indicates that DES causes disruption of the mitotic spindle, centriole elongation, and unusual chromosome associations due to interkinetochore microtubules. No apparent damage to kinetochores was noted in lagging or maloriented chromosomes.