Analysis of the distribution of the kinetochore protein Ndc10p in Saccharomyces cerevisiae using 3-D modeling of mitotic spindles.
Ndc10p is one of the DNA-binding constituents of the kinetochore in Saccharomyces cerevisiae but light microscopy analysis suggests that Ndc10p is not limited to kinetochore regions. We examined the localization of Ndc10p using immunoelectron microscopy and showed that Ndc10p is associated with spindle microtubules from S-phase through anaphase. By serial section reconstruction of mitotic spindles combined with immunogold detection, we showed that Ndc10p interacts with microtubules laterally as well as terminally. About 50% of the gold label in serial section reconstructions of short mitotic spindles was associated with the walls of spindle microtubules. Interaction of kinetochore components with microtubule walls was also shown for kinetochore protein Ndc80p. Our data suggest that at least a subset of kinetochore-associated protein is dispersed throughout the mitotic spindle.
Müller-Reichert
Thomas
T
Max Planck Institute of Molecular Cell Biology and Genetics, Pfotenhauerstrasse 108, 01307 Dresden, Germany. mueller-reichert@mpi-cbg.de
Sassoon
Ingrid
I
O'Toole
Eileen
E
Romao
Maryse
M
Ashford
Anthony J
AJ
Hyman
Anthony A
AA
Antony
Claude
C
D000705:Anaphase; D002503:Centromere / Q000648:ultrastructure; D004268:DNA-Binding Proteins / Q000096:biosynthesis* / Q000737:chemistry; D007150:Immunohistochemistry; D018386:Kinetochores / Q000378:metabolism*; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000737:chemistry; D011485:Protein Binding; D016196:S Phase; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000737:chemistry; D008941:Spindle Apparatus
C081071:CBF2 protein, S cerevisiae; D004268:DNA-Binding Proteins; C112472:NDC80 protein, S cerevisiae; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins