Developmental lineage priming in Dictyostelium by heterogeneous Ras activation
- University of Manchester, United Kingdom
- University of British Columbia, Canada
Figures
Figure 1 with 3 supplements
RasGEFE mutant cells are enriched in a genetic screen for modulators of nutritional bias.
(A) G− cells produce fewer spores than G+ cells in chimeric development. GFP-labelled Ax3 wild type cells were grown in either G+ or G− conditions and mixed 10:90 with wild type G+ cells. GFP spores …
Figure 1—figure supplement 1
Organisation of cell types in Dictyostelim slug and culminants.
Nomenclatures for parts of Dictyostelium slug (left) and fruiting body (right) are depicted. Yellow (prespore) cells eventually form sorus and red cells (pstA or AB) form stalk cells. Blue (pstO) …
Figure 1—figure supplement 2
Cell type specific effects of nutritional history in Dictyostelium slugs.
(A) and culminants (B) 10% labelled G+ cells (upper panels) or G− cells (lower panels) mixed with 90% unlabelled G+ or G− cells. Closed arrows indicate enrichment of label, open arrows an absence. …
Figure 1—figure supplement 3
Enrichment of REMI mutants during screen correlates with change in cell fate preference during development.
(A) Patterning phenotype of REMI mutant pools in chimera with wild type cells. 10% wild type GFP cells were mixed with 90% REMI mutant cells from each pool at each round of selection. The dark …
Figure 2 with 2 supplements
gefE− mutant cells avoid pstO and pstB fates.
(A) RFP-labelled AX3 wild type cells were mixed at a 50:50 ratio with unlabelled wild type or gefE− mutant cells. Both strains were grown in presence of glucose. Number of unlabelled spores …
Figure 2—figure supplement 1
Cell type specific effects in chimeric gefE−/wt slugs.
(A) and culminants (B) 10% labelled Ax3 wild type (upper panels) or gefE− mutant cells (lower panels) mixed with 90% unlabelled wild type or gefE− mutant cells. Closed arrows indicate enrichment of …
Figure 2—figure supplement 2
gefE− mutant cells avoid pstO and pstB fates when both partners have a G- growth history.
All cells were grown in absence of glucose. 10% GFP-labelled Ax3 wild type or gefE− mutant cells were then mixed with 90% wild type or gefE− mutant cells and developed to slug stage.
Figure 3
Cell type differentiation is unaffected during clonal development in the gefE− mutant.
(A) Quantification of prespore:prestalk ratio and total number of spores produced by Ax3 wild type and gefE− mutant at slug and fruiting body stages respectively. (B) Expression of prestalk markers e…
Figure 4
gefE− mutant cells are less sensitive to DIF.
(A) Quantification of stalk cell formation 22 hr after DIF induction. (B) Expression level of lacZ reporter gene fused to prestalk specific promoter of ecmA (left) or ecmB (right) 22 hr after DIF …
Figure 5
GefE regulates the DIF response threshold.
(A) Replacement of the endogenous ecmA gene with GFP (B) prestalk specific expression of GFP knock-in strain at slug and culminant stages. (C) FACS analysis of Ax3 wild type and gefE− mutant GFP …
Figure 6 with 1 supplement
GefE activates RasD.
(A) Comparison of the levels of activated RasD-GTP and total RasD by Western blot in vegetative or 12 hr starved cells grown in the presence or absence of glucose. (B) Comparison of activated …
Figure 6—figure supplement 1
rasD− mutant cells avoid pstO and pstB fates in chimera with wild type cells.
10% Ax3 labelled wild type were mixed with 90% unlabelled rasD− mutant cells. The top row shows the control mix and is comparable to the image directly beneath. Closed arrows indicate enrichment of …
Figure 7 with 1 supplement
RasD expression is heterogeneous in growth phase populations.
(A) Dual promoter vectors used to drive constitutive GFP expression and gefE promoter (left) or rasD promoter (right) driven RFP expression. Cells growing in tissue culture plates were photographed …
Figure 7—figure supplement 1
RasD is expressed in prestalk cells during development.
Expression of rasD:RFP (‘text’) in the prestalk populations of a clonal slug (left) and culminant (right). AP axis of both structures oriented top-bottom.
