Components of the yeast spindle and spindle pole body.
Yeast spindle pole bodies (SPBs) with attached nuclear microtubles were enriched approximately 600-fold from yeast cell extracts. 14 mAbs prepared against this enriched SPB fraction define at least three components of the SPB and spindle. Immunofluorescent staining of yeast cells showed that throughout the cell cycle two of the components (110 and 90 kD) were localized exclusively to the SPB region, and the other (80 kD) was localized both to the SPB region and to particulate dots in short spindles. Immunoelectron microscopy confirmed and extended most of these findings. Thus the 110-kD component was localized to a layer in the SPB just to the nuclear side of the plane of the inner nuclear membrane. The 90-kD component was localized in a layer across the cytoplasmic face of intact SPBs, and, in SPBs where nuclear microtubules were removed by extraction with DEAE-dextran, the 90-kD component was also found in an inner nuclear layer close to where spindle microtubules emerge. In intact SPBs with attached nuclear microtubules the anit-80-kD mAb labels microtubules, particularly those close to the SPB. These results begin to provide a preliminary molecular map of the SPB and should also enable the corresponding genes to be isolated.
Rout
M P
MP
Medical Research Council Laboratory of Molecular Biology, Cambridge, England.
Kilmartin
J V
JV
D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D001921:Brain; D002417:Cattle; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000302:isolation & purification*; D015151:Immunoblotting; D008722:Methods; D008856:Microscopy, Fluorescence; D016253:Microscopy, Immunoelectron; D008870:Microtubules / Q000378:metabolism; D008954:Models, Biological; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000648:ultrastructure; D008941:Spindle Apparatus / Q000737:chemistry* / Q000502:physiology / Q000648:ultrastructure
D000911:Antibodies, Monoclonal; D005656:Fungal Proteins