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HP1 proteins are conserved non histone chromosomal proteins involved in the epigenetic repression of transcription. Three HP1 proteins, HP1alpha, HP1beta and HP1gamma are expressed in mammalian cells. Polyclonal antibodies directed against peptides specific for HP1alpha and HP1gamma were elicited in rabbits, affinity purified, then used to localize both proteins on spreads of unfixed metaphasic chromosomes. We show here, by conventional and confocal microscopy, that both proteins are localized at centromeres and pericentromeres, and that HP1gamma is also present on euchromatic sites of chromosome arms. Show less

Mammalian heterochromatin proteins 1 (HP1alpha, HP1beta, and HP1gamma) are nonhistone proteins that interact in vitro with a set of proteins that play a role in chromatin silencing, transcription, and chromatin remodeling. Using antibodies specific for each HP1 isoform, we showed that they segregate in distinct nuclear domains of human HeLa cells. By contrast, in mouse 3T3 interphase cells, HP1alpha and HP1beta are strictly colocalized. In mitotic HeLa cells, all of HP1alpha and a fraction of HP1beta and HP1gamma remain associated with chromosomes. Immunostaining of spread HeLa chromosomes showed that HP1alpha is mainly localized on centromeres as shown previously for HP1beta, while HP1gamma is distributed on discrete sites on the arms of chromosomes. Biochemical analysis showed that HP1alpha and HP1gamma are phosphorylated throughout the cell cycle, although more extensively in mitosis than in interphase, while HP1beta apparently remains unphosphorylated. Therefore, despite their extensive sequence conservation, mammalian HP1 isoforms differ widely in their nuclear localization, mitotic distribution and cell cycle-related phosphorylation. Thus, subtle differences in primary sequence and in posttranslational modifications may promote their targeting at different chromatin sites, generating pleiotropic effects. Show less