Dissection of fission yeast microtubule associating protein p93Dis1: regions implicated in regulated localization and microtubule interaction.
BACKGROUND
Fission yeast microtubule associating protein (MAP) p93Dis1 functions for sister chromatid separation: dis1 mutants fail to separate chromosomes, while the spindle elongates but without cyclin destruction. p93Dis1 localizes along microtubules in interphase cytoplasm, but shifts to the spindle pole body (SPB) and spindle microtubules upon the entry into mitosis. In this study, regions of p93Dis1 were dissected to examine their role.


RESULTS
Nitrocellulose filter blotting shows that recombinant Dis1 binds to bovine brain microtubules in vitro. A basic central region rich in S, T and P is essential for this association. However, the whole p93Dis1 with N- and C-termini containing a conserved repeat motif and heptad repeats, respectively, is necessary for normal microtubule association in vivo. The N-truncated region also binds to microtubules but only to the portions near the SPBs. Overproduction phenotypes indicate that p93Dis1 greatly affects spindle formation and cell morphogenesis. The central region is essential but, by itself, not sufficient for generating such effects.


CONCLUSIONS
We propose that p93Dis1 consists of three regions which carry distinct properties for localization: the N-region for cell cycle dependent localization, the central region for direct microtubule association, and the C-region for SPB and nuclear localization. The essential role of p93Dis1 is carried out in the C-region, while the N-region acts as a regulator.
Nakaseko
Y
Y
Department of Biophysics, Graduate School of Science, Kyoto University, Japan.
Nabeshima
K
K
Kinoshita
K
K
Yanagida
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism; D002417:Cattle; D002453:Cell Cycle; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D066298:In Vitro Techniques; D008869:Microtubule-Associated Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D008938:Mitosis; D008954:Models, Biological; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011485:Protein Binding; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins; D008941:Spindle Apparatus / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D005656:Fungal Proteins; D008869:Microtubule-Associated Proteins; D029702:Schizosaccharomyces pombe Proteins; C094879:dis1 protein, S pombe