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© The Rockefeller University Press, 0021-9525/1997//229 $5.00
The Journal of Cell Biology, Volume 136, Number 2, , 1997 229-240

Article

Chromosome Fragments Possessing Only One Kinetochore Can Congress to the Spindle Equator



Alexey Khodjakov*, Richard W. Cole*, Bruce F. McEwen*,{ddagger}, Karolyn F. Buttle*, and Conly L. Rieder{ddagger},*

* Division of Molecular Medicine, Wadsworth Center, New York State Department of Health, Albany, New York 12201-0509; and {ddagger} Department of Biomedical Sciences, State University of New York, Albany, New York 12201

We used laser microsurgery to cut between the two sister kinetochores on bioriented prometaphase chromosomes to produce two chromosome fragments containing one kinetochore (CF1K). Each of these CF1Ks then always moved toward the spindle pole to which their kinetochores were attached before initiating the poleward and away-from-the-pole oscillatory motions characteristic of monooriented chromosomes. CF1Ks then either: (a) remained closely associated with this pole until anaphase (50%), (b) moved (i.e., congressed) to the spindle equator (38%), where they usually (13/19 cells) remained stably positioned throughout the ensuing anaphase, or (c) reoriented and moved to the other pole (12%). Behavior of congressing CF1Ks was indistinguishable from that of congressing chromosomes containing two sister kinetochores. Three-dimensional electron microscopic tomographic reconstructions of CF1Ks stably positioned on the spindle equator during anaphase revealed that the single kinetochore was highly stretched and/or fragmented and that numerous microtubules derived from the opposing spindle poles terminated in its structure. These observations reveal that a single kinetochore is capable of simultaneously supporting the function of two sister kinetochores during chromosome congression and imply that vertebrate kinetochores consist of multiple domains whose motility states can be regulated independently.

Abbreviations used in this paper: AP, away-from-the-pole; CF1K, chromosome fragment containing one kinetochore; IVEM, intermediate voltage electron microscope; K-fiber, kinetochore fiber; Mt, microtubule; MUG, mitosis with unreplicating genome; P, poleward.

Address all correspondence to Dr. Conly L. Rieder, Division of Molecular Medicine, Wadsworth Center, PO. Box 509, Albany, NY 12201-0509. Tel.: (518) 474-6774. Fax: (518) 486-4901. E-mail: RIEDER@ WADSWORTH. ORG


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