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First published online 2 July 2003
doi: 10.1242/jcs.00635
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Research Article |
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
, ß, and 
, 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 and were more
preferentially localized to PML nuclear bodies. During metaphase, only
HP1, 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 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 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 fragment
accumulated in centromeric regions. These results indicate that different
specific regions of human HP1 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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