Stu1p is physically associated with beta-tubulin and is required for structural integrity of the mitotic spindle.
Formation of the bipolar mitotic spindle relies on a balance of forces acting on the spindle poles. The primary outward force is generated by the kinesin-related proteins of the BimC family that cross-link antiparallel interpolar microtubules and slide them past each other. Here, we provide evidence that Stu1p is also required for the production of this outward force in the yeast Saccharomyces cerevisiae. In the temperature-sensitive stu1-5 mutant, spindle pole separation is inhibited, and preanaphase spindles collapse, with their previously separated poles being drawn together. The temperature sensitivity of stu1-5 can be suppressed by doubling the dosage of Cin8p, a yeast BimC kinesin-related protein. Stu1p was observed to be a component of the mitotic spindle localizing to the midregion of anaphase spindles. It also binds to microtubules in vitro, and we have examined the nature of this interaction. We show that Stu1p interacts specifically with beta-tubulin and identify the domains required for this interaction on both Stu1p and beta-tubulin. Taken together, these findings suggest that Stu1p binds to interpolar microtubules of the mitotic spindle and plays an essential role in their ability to provide an outward force on the spindle poles.
Yin
Hongwei
H
Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853-2703, USA.
You
Liru
L
Pasqualone
Danielle
D
Kopski
Kristen M
KM
Huffaker
Tim C
TC
D001483:Base Sequence; D017931:DNA Primers; D005838:Genotype; D008869:Microtubule-Associated Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008958:Models, Molecular; D016296:Mutagenesis; D014176:Protein Biosynthesis; D011487:Protein Conformation; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008941:Spindle Apparatus / Q000502:physiology / Q000648:ultrastructure*; D014158:Transcription, Genetic; D014404:Tubulin / Q000378:metabolism*
D017931:DNA Primers; D008869:Microtubule-Associated Proteins; D012097:Repressor Proteins; C091278:STU1 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D014404:Tubulin