Fig. 5. Transgene deletions demonstrate that a functional GAP domain is required for Tum cytokinetic function. Tum transgenes with GAP domain deletions UAS-tum
EIE (A-E) and UAS-tum
YRL (F-L), were expressed in prd stripes in tumDH15 embryos. In all panels, anti-myc antibody shows the transgene localisation in red, tubulin (or F-actin, panel F) localisation is shown in green and DNA in blue. (A-E) When UAS-tum
EIE is expressed in tumDH15 embryos, it fails to localise correctly or rescue cytokinesis. Binucleate cells (asterisks, A) are readily detected in stripes of UAS-tum
EIE-expressing cells, whereas midbody remnants are rarely found. Cells in the mitotic cycle in these stripes have diffuse Tum
EIE protein at metaphase (arrow, C), anaphase (arrowhead, C) and telophase (arrow, E). (F-J) When UAS-tum
YRL is expressed in tumDH15 embryos, it localises to the cortex equator, but also fails to rescue cytokinesis. Binucleate cells (asterisks, F) are readily detected in stripes of UAS-tum
YRL-expressing cells, whereas midbody remnants are rarely found. Tum
YRL localises to the ends of microtubules at the cell equator at anaphase (arrowhead, F) and this localisation does not change throughout telophase or the subsequent interphase (arrowheads, G,H,I,J). Bar, 5 µm. Grayscale images for all color channels can be viewed in supplementary material Fig. S4.