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© The Rockefeller University Press, 0021-9525/2000//531 $5.00
The Journal of Cell Biology, Volume 149, Number 3, , 2000 531-536

A Kinase–Anchoring Protein (Akap95) Recruits Human Chromosome-Associated Protein (Hcap-D2/Eg7) for Chromosome Condensation in Mitotic Extract

^a Institute of Medical Biochemistry, Faculty of Medicine, University of Oslo, 0317 Oslo, Norway
^b CNRS UPR41, Biologie et Génétique du Développement, Faculté de Médecine, 35043 Rennes Cedex, France
Institute of Medical Biochemistry, Faculty of Medicine, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway.+47-22851497+47-22851013

philippe.collas{at}basalmed.uio.no

Association of the condensin multiprotein complex with chromatin is required for chromosome condensation at mitosis. What regulates condensin targeting to chromatin is largely unknown. We previously showed that the nuclear A kinase–anchoring protein, AKAP95, is implicated in chromosome condensation. We demonstrate here that AKAP95 acts as a targeting protein for human chromosome-associated protein (hCAP)-D2/Eg7, a component of the human condensin complex, to chromosomes. In HeLa cell mitotic extract, AKAP95 redistributes from the nuclear matrix to chromatin. When association of AKAP95 with chromatin is prevented, the chromatin does not condense. Condensation is rescued by a recombinant AKAP95 peptide containing the 306 COOH-terminal amino acids of AKAP95. Recombinant AKAP95 binds chromatin and elicits recruitment of Eg7 to chromosomes in a concentration-dependent manner. Amount of Eg7 recruited correlates with extent of chromosome condensation: resolution into distinct chromosomes is obtained only when near-endogenous levels of Eg7 are recruited. Eg7 and AKAP95 immunofluorescently colocalize to the central region of methanol-fixed metaphase chromosomes. GST pull-down data also suggest that AKAP95 recruits several condensin subunits. The results implicate AKAP95 as a receptor that assists condensin targeting to chromosomes.

Key Words: chromosome • condensin • AKAP • hCAP • mitosis

© 2000 The Rockefeller University Press

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