Cell differentiation in a temperature-sensitive stalkless mutant of Dictyostelium discoideum

Development ◽  
1983 ◽  
Vol 74 (1) ◽  
pp. 235-243
Author(s):  
Aiko Amagai ◽  
Shuji Ishida ◽  
Ikuo Takeuchi

A temperature-sensitive aggregateless and stalkless mutant was isolated from Dictyostelium discoideum NC-4. The mutant cells cannot aggregate at 27°C, but aggregate and form normal fruiting bodies at 21°C. When the temperature was shifted to 27°C after aggregation at 21°C, almost all of the cells in the aggregate differentiated into spores. Neither stalk cells nor stalk tubes formed at 27°C. Inhibition of stalk formation was not lifted by addition of cyclic AMP. Nevertheless, the proportion of prespore to total cells within the mutant slugs was normal, at both 21 °C and 27 °C. At 27 °C, a slug was transformed into a spherical cell mass at the end of migration, within which pre-existing prespore cells differentiated into spores. The remaining prestalk cells were then converted to prespore cells which later became spores. As the cell-type conversion continued, formation of a spore mass resulted. The development of the mutant is thus consistent with the idea that the presumptive cell differentiation is directly related to the terminal cell differentiation. During migration at 27 °C, the number of prestalk cells decreased in the anterior part of the slug but instead increased at the foot or the rear part, whereas the prestalk—prespore pattern remained normal at 21 °C. The fact that a normal proportion of prespore cells was maintained in spite of their deranged distribution at 27 °C indicates that the regulation of proportion is independent of the formation of pattern.

1984 ◽  
Vol 4 (6) ◽  
pp. 1035-1041 ◽  
Author(s):  
S Biswas ◽  
S C Kayman ◽  
M Clarke

Dictyostelium discoideum MC2 is a temperature-sensitive motility mutant of AX3. Mutant cells are incapable of growth, phagocytosis, and migration under restrictive conditions (Kayman et al., J. Cell Biol. 92:705-711, 1982). We show here that at the restrictive temperature MC2 cells grown axenically or on bacteria synthesized excessive quantities of the lectin discoidin I. By two-dimensional gel electrophoresis and peptide mapping, the proteins overproduced by MC2 cells were indistinguishable from discoidin I synthesized at lower levels in AX3 cells. At least two of the three species of discoidin I were overproduced. This protein family constituted 9% of the total protein in cells that were incubated overnight at 27 degrees C in axenic medium. Although MC2 cells are defective in nutrient uptake under restrictive conditions, the overproduction of discoidin I did not appear to be part of a pleiotropic response to starvation. We propose that transcription of the coordinately regulated discoidin I genes is altered in mutant cells. This alteration may be related to the motility defects manifested by MC2.


2004 ◽  
Vol 3 (5) ◽  
pp. 1241-1248 ◽  
Author(s):  
Toshinari Maruo ◽  
Haruyo Sakamoto ◽  
Negin Iranfar ◽  
Danny Fuller ◽  
Takahiro Morio ◽  
...  

ABSTRACT We have determined the proportions of the prespore and prestalk regions in Dictyostelium discoideum slugs by in situ hybridization with a large number of prespore- and prestalk-specific genes. Microarrays were used to discover genes expressed in a cell type-specific manner. Fifty-four prespore-specific genes were verified by in situ hybridization, including 18 that had been previously shown to be cell type specific. The 36 new genes more than doubles the number of available prespore markers. At the slug stage, the prespore genes hybridized to cells uniformly in the posterior 80% of wild-type slugs but hybridized to the posterior 90% of slugs lacking the secreted alkylphenone differentiation-inducing factor 1 (DIF-1). There was a compensatory twofold decrease in prestalk cells in DIF-less slugs. Removal of prespore cells resulted in cell type conversion in both wild-type and DIF-less anterior fragments. Thus, DIF-1 appears to act in concert with other processes to establish cell type proportions.


1984 ◽  
Vol 4 (6) ◽  
pp. 1035-1041
Author(s):  
S Biswas ◽  
S C Kayman ◽  
M Clarke

Dictyostelium discoideum MC2 is a temperature-sensitive motility mutant of AX3. Mutant cells are incapable of growth, phagocytosis, and migration under restrictive conditions (Kayman et al., J. Cell Biol. 92:705-711, 1982). We show here that at the restrictive temperature MC2 cells grown axenically or on bacteria synthesized excessive quantities of the lectin discoidin I. By two-dimensional gel electrophoresis and peptide mapping, the proteins overproduced by MC2 cells were indistinguishable from discoidin I synthesized at lower levels in AX3 cells. At least two of the three species of discoidin I were overproduced. This protein family constituted 9% of the total protein in cells that were incubated overnight at 27 degrees C in axenic medium. Although MC2 cells are defective in nutrient uptake under restrictive conditions, the overproduction of discoidin I did not appear to be part of a pleiotropic response to starvation. We propose that transcription of the coordinately regulated discoidin I genes is altered in mutant cells. This alteration may be related to the motility defects manifested by MC2.


1995 ◽  
Vol 128 (3) ◽  
pp. 405-413 ◽  
Author(s):  
J E Segall ◽  
A Kuspa ◽  
G Shaulsky ◽  
M Ecke ◽  
M Maeda ◽  
...  

Analysis of a developmental mutant in Dictyostelium discoideum which is unable to initiate morphogenesis has shown that a protein kinase of the MAP kinase/ERK family affects relay of the cAMP chemotactic signal and cell differentiation. Strains in which the locus encoding ERK2 is disrupted respond to a pulse of cAMP by synthesizing cGMP normally but show little synthesis of cAMP. Since mutant cells lacking ERK2 contain normal levels of both the cytosolic regulator of adenylyl cyclase (CRAC) and manganese-activatable adenylyl cyclase, it appears that this kinase is important for receptor-mediated activation of adenylyl cyclase.


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