[x] Longer cdc13 signal, this is clear evidence (Fig. 3)
[x] Intranuclear astral microtubules (Fig. 2)
Fig. 3
[x] pdk1D (background cdc25-22)
[x] Delay in ring assembly -> not only caused by delay in anaphase onset (see below)
[x] 31% kinetochores associated with mad2 vs. 20% wt (metaphase delay may be caused by kinetochore attachment problems)
[x] pdk1D mph1D (background cdc25-22)
[x] anaphase delay is rescued (no checkpoint) -> genetic interaction phenotypic suppression
[x] delay in ring assembly (not fully rescued)
Fig. 4
[x] Pdk1 -> Localizes to the SPB during mitosis only, lost at late anaphase. Also to the medial-cortical region weakly, even in pressence of latrunculin.
Fig. 5
[x] pdk1D + plo1-1 and plo1-25 + pdk1D AT COLD TEMP
manulera
commented
2 years ago
PMID:15857958
Roles of Pdk1p, a Fission Yeast Protein Related to Phosphoinositide-dependent Protein Kinase, in the Regulation of Mitosis and Cytokinesis
Proteins related to the phosphoinositide-dependent protein kinase family have been identified in the majority of eukaryotes. Although much is known about upstream mechanisms that regulate the PDK1-family of kinases in metazoans, how these kinases regulate cell growth and division remains unclear. Here, we characterize a fission yeast protein related to members of this family, which we have termed Pdk1p. Pdk1p localizes to the spindle pole body and the actomyosin ring in early mitotic cells. Cells deleted for pdk1 display multiple defects in mitosis and cytokinesis, all of which are exacerbated when the function of fission yeast polo kinase, Plo1p, is partially compromised. We conclude that Pdk1p functions in concert with Plo1p to regulate multiple processes such as the establishment of a bipolar mitotic spindle, transition to anaphase, placement of the actomyosin ring and proper execution of cytokinesis. We also present evidence that the effects of Pdk1p on cytokinesis are likely mediated via the fission yeast anillin-related protein, Mid1p, and the septation initiation network. © 2005 by The American Society for Cell Biology.
Results
Fig. 2
Fig. 3
[x] pdk1D (background cdc25-22)
[x] pdk1D mph1D (background cdc25-22)
Fig. 4
Fig. 5
Fig. 5
At normal temp
Fig. 6
Fig. 7
[x] pdk1D
[x] mid1D cells don't localize pdk1D to the cortical center (site of cell division) -> depends on annotation!
[x] pdk1D sid2-250, pdk1D spg1-106, pdk1D sid4-A1 were unable to divide and form colonies at all