FIGURE 6:

Overproduction of Alp1^D causes disruption of microtubule structure, which can be rescued with co-overproduction of GDP-bound Alp41. (A) Cells transformed with an overexpression plasmid of Alp1^D were cultured under minus-thiamine conditions for 20 h. Note that cells have not yet undergone complete microtubule disruption, by which colocalization of Alp1^D with the remaining microtubules is visible (marked with arrowheads). (B) Single-cell images in which there is clear localization of the GFP-Alp1^D on the microtubules. (C) Cells were double transformed with overexpression plasmids of Alp1^D and a wild-type, GTP-bound (Q70L), or GDP-bound (T30N) form of Alp41. Green arrowheads indicate cells that show GFP-Alp1^D colocalizing with microtubules. Red arrowheads indicate cells where this colocalization is not seen. (D) Quantification of GFP-Alp1^D localization. Two hundred cells showing GFP-Alp1^D overproduction were counted and categorized into cells showing colocalization of GFP-Alp1^D signal on microtubules or cells showing no colocalization. Bars, 5 μm.