Characterization of a Novel Kinetochore Protein, CENP-H*
- From the ‡Tsukuba Life Science Center, The Institute of Physical and Chemical Research (RIKEN), 3-1, Koyadai, Tsukuba, Ibaraki 305-0074, Japan and the §Institute of Applied Biochemistry, Tsukuba University, 1-1, Tennoudai, Tsukuba, Ibaraki 305-8572, Japan
Abstract
Macromolecular centromere-kinetochore complex plays a critical role in sister chromatid separation, but its complete protein composition as well as its precise dynamic function during mitosis has not yet been clearly determined. Here we report the isolation of a novel mouse kinetochore protein, CENP-H. The CENP-H, with an apparent molecular mass of 33 kDa, was found to contain a coiled-coil structure and a nuclear localization signal. The CENP-H transcripts were relatively scarce but were detectable in most tissues and embryos at various stages of development. Immunofluorescence stainings of mouse fibroblast cells with anti-CENP-H-specific antibody demonstrated that the CENP-H is specifically and constitutively localized in kinetochores throughout the cell cycle; this was also confirmed by stainings with anti-centromere-specific antibody. Thus the newly isolated CENP-H may play a role in kinetochore organization and function throughout the cell cycle.
Footnotes
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↵* This work was supported in part by a Special Grant for Promotion of Research from the Institute of Physical and Chemical Research (RIKEN) and by grants from the Ministry of Education, Science and Culture of Japan and the Science and Technology Agency of Japan.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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↵¶ To whom correspondence should be addressed. Tel.: +81 298 36 9075; Fax: +81 298 36 9090; E-mail: todokoro@rtc.riken. go.jp.
- Abbreviations:
- MCAK
-
mitotic centromere-associated kinesin
- bp
-
base pair(s)
- 5′-RACE
-
5′ rapid amplification of cDNA ends
- PCR
-
polymerase chain reaction
- PIPES
-
1,4-piperazinediethanesulfonic acid
- PBS
-
phosphate-buffered saline
- FITC
-
fluorescein isothiocyanate
- DAPI
-
4,6-diamidino-2-phenylindole
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- Received June 28, 1999.
- Revision received July 26, 1999.
- The American Society for Biochemistry and Molecular Biology, Inc.
