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First published online 2 July 2003
doi: 10.1242/jcs.00635


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Journal of Cell Science 116, 3327-3338 (2003)
doi: 10.1242/jcs.00635


Research Article

Cell cycle behavior of human HP1 subtypes: distinct molecular domains of HP1 are required for their centromeric localization during interphase and metaphase

Tomohiro Hayakawa1,2, Tokuko Haraguchi1,2, Hiroshi Masumoto3 and Yasushi Hiraoka1,2,*

1 CREST Research Project, Kansai Advanced Research Center, Communications Research Laboratory, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe 651-2492, Japan
2 Department of Biology, Graduate School of Science, Osaka University,1-1 Machikaneyama, Osaka 560-0043, Japan
3 Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan

* Author for correspondence (e-mail: yasushi{at}crl.go.jp)

Accepted 28 April 2003

Heterochromatin protein 1 (HP1) plays an important role in heterochromatin formation. Three subtypes of HP1, namely HP1{alpha}, ß, and {gamma}, have been identified in humans. In this study, using yellow fluorescent protein (YFP) fusion constructs, we examined the intracellular localization of human HP1 subtypes during the cell cycle. During interphase, all three HP1 subtypes were localized to centromeric heterochromatin and to promyelocytic leukemia (PML) nuclear bodies. Different preferences, however, were observed among the subtypes: during interphase HP1ß localized most preferentially to centromeric heterochromatin, whereas HP1{alpha} and {gamma} were more preferentially localized to PML nuclear bodies. During metaphase, only HP1{alpha}, was localized to the centromere. We thus determined which molecular domains of HP1 were necessary for their intracellular localization. Our results showed that the C-terminal fragment (amino acid residues 101-180) of HP1{alpha} was necessary for localization to the metaphase centromere and the N-terminal fragment (amino acid residues 1-76) of HP1ß was necessary for localization to the interphase centromere. Interestingly, simultaneous observations of residues 101-180 of HP1{alpha} and residues 1-76 of HP1ß in living HeLa cells revealed that during late prophase, the HP1ß fragment dissociated from centromeric regions and the HP1{alpha} fragment accumulated in centromeric regions. These results indicate that different specific regions of human HP1{alpha} and HP1ß mediate localization to metaphase and interphase centromeric regions resulting in association of different subtypes of HP1 with the centromere at different times during the cell cycle.

Key words: Heterochromatin, HP1, PML, Centromere, CENP-B




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