JCS02501 Supplementary Material

Files in this Data Supplement:

• Movie 1 - Movie 1. Nuclear migration in wild type. In wild type, nuclei migrate along hyphae and into new branches. They maintain even spacing. Nuclei are labelled with GFP. During mitosis, nuclei are not visible, because the GFP construct is excluded from the nucleus and localizes to the cytoplasm. Strain is SPR50.

• Movie 2 - Movie 2. Nuclear migration in apsB mutant. In an apsB6 strain, nuclei migrate independently of each other in a chaotic manner. The arrow follows a nucleus passing over another nucleus (arrow head). Nuclei are not spaced evenly and show chaoic movement into and out of branches. Nuclei are labelled with GFP. During mitosis nuclei are not visible, because the GFP construct is excluded from the nucleus and localizes to the cytoplasm. Strain is SRS11.

• Movie 3 - Movie 3. Nuclear navigation in apsB mutants. Similar to Movie 2, but nuclear navigation into branching hyphae can be followed easily due to coloration of individual nuclei. Strain is SRS11.

• Movie 4 - Movie 4. Nuclear movement in apsA mutants. Nuclear movement in the apsA1 mutant strain. Nuclei are clustered and show chaotic movement. Nuclei are labelled with GFP. Strain is SDV12.

• Movie 5 - Movie 5. ApsA at the cortex. Localization of ApsA-GFP at the cortex of a fresly germinated spore. Confocal laser scanning image. Strain is SSN18.

• Movie 6 - Movie 6 (and Fig. 1C). Spindle movement in wild type. Mitotic spindles oscillate in wild type. MTs were stained with GFP. Strain is SJW02.

• Movie 7 - Movie 7 (Fig. 1D). Spindle movement in apsA mutants. In the DapsA strain, miotic spindles never moved. MTs were stained with GFP. Strain is SNS9.

• Movie 8 - Movie 8 (Fig. 2). Movement of ApsB along MTs. Arrows follow ApsB-GFP spots. ApsB-GFP moves bidirectional along MTs (stained with GFP). Notice that the spot in the left cell rotates around the MT axis. Strain is SDM40.

• Movie 9 - Movie 9 (Fig. 2). Movement of ApsB. ApsB-GFP moves very fast (up to 6 µm/seconds) along MTs in both directions. Strain is SDM40.

• Movie 10 - Movie 10.MT bundles and ApsB. Nuclei, MTs and ApsB are labelled with GFP. At the beginning a predominant, thick MT (bundle) can be seen, to which four nuclei (big white ‘eggs’) are connected with their spindle pole bodies (ApsB-GFP, small bright dots). The asterisk indicates the event of a nucleus separating from the ‘mother MT bundle’, while still being connected to a thinner MT filament and migrating a few microns. The arrow follows an ApsB-GFP spot during its movement along MTs. Notice that the spot can change its track easily and that it moves into both directions. At  one time point a separated MT reassociates with the ‘mother bundle’ at the position where the ApsB-spot is ‘pausing’ (indicated by ‘X’). Strain is SDM40.

• Movie 10a - Movie 10a. MT bundles and ApsB. Scheme of ApsB movement along MT filaments. Notice the bidirectional transport and that this bidirectionality could be achieved by either two motors with different polarity or by a single motor moving along anti-parallel filaments, while easily changing the tracks.

• Movie 11 - Movie 11 (and Fig. 4B). GFP-KipA at MTOC in WT. KipA is a kinesin-like protein and can be used as a MT-plus end marker (see (Konzack et al., 2005). GFP-KipA comets emerging at septa and SPBs indicate MTOCs. A quantitative analysis of signal counts can be used as a an indicator of MTOC activity comparing the situation in WT and apsB mutants. Strain is SSK92.

• Movie 12 - Movie 12. (and Fig. 5A). Nuclear movement. MTs (labelled with GFP) emerge from the SPB of the nucleus (dark, negatively stained ball). The nucleus with its leading SPB is pulled upward. It appeares that only MTs at its front side apply the pulling force. Strain is SJW02.

• Movie 13 - Movie 13. (and Fig. 5B). Nuclear movement. Similar to Movie 12, a nucleus is moving due to a pulling force applied to its SPB. Notice that the SPB changes its position from one side (top) to the opposite side (bottom) of the nucleus, dependent on the direction of the force applied to the attached MTs. Strain is SJW02.

• Movie 14 - Movie 14. (and Fig. 5C). Synchronous movement of nuclei. Adjacent nuclei, connected to the same MT bundle, move synchronous if an appropriate force is applied. Nuclei, MTs and ApsB are labelled with GFP. Notice that, MT number is reduced due to the dominant negative influence of the C-terminal tagged ApsB-GFP. Strain is SDM40.

• Movie 15 - Movie 15. MTs connect adjacent nuclei. In wild type, movement of one nucleus can also be coordinated to the movement of a second, adjacent nucleus, if connected to the same MT (bundle or filament). MTs are labelled with GFP. Nuclei can be seen as negatively stained, dark balls. One nucleus is located in the spore and the second nucleus in the freshly germinated hypha. Notice the SPBs of the nuclei as bright, white spots with emerging MT filaments. Strain is SJW02.

• Movie 16 - Movie 16 (and Fig. 5D). Nuclear movement. Two nuclei can be seen as white balls. The lower nucleus is moving upward with its SPB at its front. Notice the tear drop shape of the nucleus and a snapping of the MT filament from the right to the left side of the hypha. The nucleus is following this movement. Interestingly, nuclei can still migrate, although MT number (especially the MTs emerging from the SPB and connecting the  nucleus to the cortex) is reduced, due to the dominat negative influence of the C-terminal tagged ApsB-GFP. Strain is SDM40.

• Movie 17 - Movie 17 (and Fig. 7). ApsB accumulation in dynein mutant strains. ApsB-GFP accumulates at hyhal tips in a DnudA strain.
• Movie 18 - Movie 18 (and Fig. 7). ApsB movement in dynein mutant strains. Although ApsB-GFP accumulates at hyhal tips in a DnudA strain, it is still highly mobile. Strain is SDM92.