A Dominant Mutant of Inner Centromere Protein (INCENP), a Chromosomal Protein, Disrupts Prometaphase Congression and Cytokinesis

INCENP is a tightly bound chromosomal protein that transfers to the spindle midzone at the metaphase/anaphase transition. Here, we show that an INCENP truncation mutant ( INCENP382-839) associates with microtubules but does not bind to chromosomes, and coats the entire spindle throughout mitosis. Fu...

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Bibliographic Details
Published in:The Journal of cell biology 1998-03, Vol.140 (5), p.991-1002
Main Authors: Mackay, Alastair M., Ainsztein, Alexandra M., Eckley, D. Mark, Earnshaw, William C.
Format: Article
Language:English
Subjects:
Anaphase
Animals
Binding Sites
Cell Division
Cell Line
Cells
Cellular biology
Centromere - metabolism
Centromeres
Chromosomal Proteins, Non-Histone - genetics
Chromosomal Proteins, Non-Histone - metabolism
Chromosomes
Cytokinesis
Cytoskeletal Proteins - genetics
Cytoskeletal Proteins - metabolism
Genes
HeLa cells
Metaphase - physiology
Microtubules
Mitosis
Mitotic spindle apparatus
Mutation
Proteins
Rabbits
Somatic cells
Spindle Apparatus
Structure-Activity Relationship
Swine
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Summary:INCENP is a tightly bound chromosomal protein that transfers to the spindle midzone at the metaphase/anaphase transition. Here, we show that an INCENP truncation mutant ( INCENP382-839) associates with microtubules but does not bind to chromosomes, and coats the entire spindle throughout mitosis. Furthermore, an INCENP truncation mutant ( INCENP43-839) previously shown not to transfer to the spindle at anaphase (Mackay, A.M., D.M. Eckley, C. Chue, and W.C. Earnshaw. 1993. J. Cell Biol. 123:373-385), is shown here to bind chromosomes, but is unable to target to the centromere. Thus, association with the chromosomes, and specifically with centromeres, appears to be essential for INCENP targeting to the correct spindle subdomain at anaphase. An INCENP truncation mutant ( INCENP1-405) that targets to centromeres but lacks the microtubule association region acquires strong dominant-negative characteristics. INCENP1-405interferes with both prometaphase chromosome alignment and the completion of cytokinesis. INCENP1-405apparently exerts its effect by displacing the endogenous protein from centromeres. These experiments provide evidence of an unexpected link between this chromosomal protein and cytokinesis, and suggest that one function of INCENP may be to integrate the chromosomal and cytoskeletal events of mitosis.
ISSN:0021-9525
1540-8140
DOI:10.1083/jcb.140.5.991