ER assembly of SNARE complexes mediating formation of partitioning membrane in Arabidopsis cytokinesis
- University of Tübingen, Germany
Figures
Figure 1 with 3 supplements
Site-specific inhibition of SNARE protein trafficking to the cell-division plane.
(A) Qa-SNARE KNOLLE trafficking route in cytokinesis (Reichardt et al., 2007). ER, endoplasmic reticulum; TGN, trans-Golgi network; CP, cell plate; MVB, multivesicular body; COPI, COPII, AP1/CCV, …
Figure 1—figure supplement 1
Cytokinetic cells in big3 and gnl1 GNL1BFA-sens. mutant seedling roots.
Mutant seedling roots were treated with 50 µM BFA for 30 min followed by 50 µM BFA + 20 µM estradiol for 210 min. Note that the population of cytokinetic cells was not altered by BFA treatment in …
Figure 1—figure supplement 2
Subcellular localisation (A, B) and co-immunoprecipitation analysis (C) of pKNOLLE::mRFP:PEP12 (aka SYP21) (red) in big3 mutant seedling root cells expressing estradiol-inducible YFP:NPSN11 (A, C) and GFP:SNAP33 (B, C).
Transgenic seedlings were treated with 50 µM BFA for 30 min followed by 50 µM BFA + 20 µM estradiol for 210 min. Protein extracts were subjected to immunoprecipitation with anti-GFP beads, protein …
Figure 1—figure supplement 3
Site-specific inhibition of SNARE protein trafficking to the cell-division plane and loss of COPI from Golgi membrane in gnl1 GNL1BFA-sens. seedlings.
Mutant seedling roots were treated with 50 µM BFA for 30 min, followed by 50 µM BFA + 20 µM estradiol for 210 min. Note cytosolic localisation of γCOP in BFA-treated cells whereas YFP:NPSN11 (Y:N11, …
Figure 2 with 1 supplement
Interaction analysis of cytokinetic SNAREs with traffic blocked at the TGN.
Wild-type (WT) and big3 mutant seedlings carrying estradiol-inducible YFP:NPSN11 (A) or GFP:SNAP33 (B) transgenes were treated with 50 µM BFA for 30 min followed by 50 µM BFA + 20 µM estradiol for …
Figure 2—figure supplement 1
Control experiments for co-immunoprecipitation analysis of cytokinetic SNAREs.
(A) No EST-induction of expression of KNOLLE SNARE partners. big3 mutant seedlings carrying YFP:NPSN11 (left) or GFP:SNAP33 (right) transgenes were treated or not treated with 50 µM BFA for 240 min. …
Figure 3
Interaction analysis of cytokinetic SNAREs with traffic blocked at the ER.
Wild-type (WT) and gnl1 mutant seedlings complemented with GNL1BFA-sens. encoding a BFA-sensitive variant of GNL1 and carrying estradiol-inducible YFP:NPSN11 (A) or GFP:SNAP33 (B) transgenes were …
Figure 4
Trafficking of cis-SNARE complexes during cytokinesis (model).
Two different types of cytokinetic cis-SNARE complexes are assembled on the ER, recruited into COPII vesicles and passed on to the Golgi stack/TGN. At the TGN, they are incorporated into AP1/CCV …
Additional files
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Supplementary file 1
(a) Frequency of cytokinetic cells in mutant seedling roots.
(b) Primers used in this study
- https://doi.org/10.7554/eLife.25327.010
