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J Cell Biol. 2015 Jun 22;209(6):813-28. doi: 10.1083/jcb.201408083.

Centromere protein F includes two sites that couple efficiently to depolymerizing microtubules.

Author information

1
Center for Theoretical Problems of Physicochemical Pharmacology, Russian Academy of Sciences, Moscow, Russia, 119991 Laboratory of Biophysics, Federal Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia, 117513 N. F. Gamaleya Research Institute for Epidemiology and Microbiology, Moscow, Russia, 123098.
2
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309.
3
Department of Bioengineering and Bioinformatics, Moscow State University, Moscow, Russia, 119991.
4
Department of Physiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104.
5
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309.
6
Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, CO 80309 richard.mcintosh@colorado.edu.

Abstract

Firm attachments between kinetochores and dynamic spindle microtubules (MTs) are important for accurate chromosome segregation. Centromere protein F (CENP-F) has been shown to include two MT-binding domains, so it may participate in this key mitotic process. Here, we show that the N-terminal MT-binding domain of CENP-F prefers curled oligomers of tubulin relative to MT walls by approximately fivefold, suggesting that it may contribute to the firm bonds between kinetochores and the flared plus ends of dynamic MTs. A polypeptide from CENP-F's C terminus also bound MTs, and either protein fragment diffused on a stable MT wall. They also followed the ends of dynamic MTs as they shortened. When either fragment was coupled to a microbead, the force it could transduce from a shortening MT averaged 3-5 pN but could exceed 10 pN, identifying CENP-F as a highly effective coupler to shortening MTs.

PMID:
26101217
PMCID:
PMC4477864
DOI:
10.1083/jcb.201408083
[Indexed for MEDLINE]
Free PMC Article

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