Application of a New Dual Localization-Affinity Purification Tag Reveals Novel Aspects of Protein Kinase Biology in Aspergillus nidulans
Figure 12
SepH displays a biphasic location to SPBs distal from the cell tip during mitosis and septation.
A Time lapse images of a germling transiting mitosis for the cell shown in Video S4. Shown is the location of SepH-DLAP together with GCP3-mCherry. SepH positive SPBs are indicated by arrowheads. In G2 over 10 non-SPB SepH foci are present in the region most distal from the cell tip (e.g. arrows, time −6′). As the cell enters prophase (p) SepH associates with tip distal SPBs and is present at SPBs in this region during metaphase (m), anaphase (a) and telophase (t). In early G1 SepH rarely locates to SPBs. The segmented line at 9 min was used to generate the kymographs in C. B The same cell in A later in G1 when 2 septa form showing that SepH again associates with SPBs distal from the cell tip in the period preceding formation of the first septa (S1) and the second septa (S2). C Kymographs generated from images captured every 90 sec from the same experiment in A and B. The position of nuclei (N1–5) is apparent from the shadows formed by the nuclear exclusion of SepH. The asterisks indicate mitotic entry when SepH enters nuclei and associates with SPBs distal from the cell tip. In early G1 few SepH foci are apparent but SepH again associates with tip distal SPBs preceding the period of septation (Sep). Note that SepH preferentially locates to SPBs on the tip distal side of where the septum subsequently forms (e.g. arrow). D Graph generated from the same experiment showing the total number of SepH foci present at each time point (green) and also indicating which of these foci correspond to SPBs (red). E DIC images from the start and end of the time course with the position of the septa indicated. Bar ∼10 µm.