INCENP centromere and spindle targeting: identification of essential conserved motifs and involvement of heterochromatin protein HP1.
The inner centromere protein (INCENP) has a modular organization, with domains required for chromosomal and cytoskeletal functions concentrated near the amino and carboxyl termini, respectively. In this study we have identified an autonomous centromere- and midbody-targeting module in the amino-terminal 68 amino acids of INCENP. Within this module, we have identified two evolutionarily conserved amino acid sequence motifs: a 13-amino acid motif that is required for targeting to centromeres and transfer to the spindle, and an 11-amino acid motif that is required for transfer to the spindle by molecules that have targeted previously to the centromere. To begin to understand the mechanisms of INCENP function in mitosis, we have performed a yeast two-hybrid screen for interacting proteins. These and subsequent in vitro binding experiments identify a physical interaction between INCENP and heterochromatin protein HP1(Hsalpha). Surprisingly, this interaction does not appear to be involved in targeting INCENP to the centromeric heterochromatin, but may instead have a role in its transfer from the chromosomes to the anaphase spindle.
Ainsztein
A M
AM
Institute of Cell and Molecular Biology, University of Edinburgh, Edinburgh EH9 3JR, Scotland, United Kingdom.
Kandels-Lewis
S E
SE
Mackay
A M
AM
Earnshaw
W C
WC
D000705:Anaphase; D000818:Animals; D001692:Biological Transport; D002460:Cell Line; D002503:Centromere / Q000378:metabolism*; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000378:metabolism*; D019143:Evolution, Molecular; D006367:HeLa Cells; D006570:Heterochromatin / Q000378:metabolism; D006801:Humans; D007668:Kidney; D046911:Macromolecular Substances; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D008941:Spindle Apparatus / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013552:Swine; D013692:Telophase; D014162:Transfection
D002868:Chromosomal Proteins, Non-Histone; D006570:Heterochromatin; C083767:INCENP protein, human; D046911:Macromolecular Substances; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D000090266:Chromobox Protein Homolog 5