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c-di-GMP induction of Dictyostelium cell death requires the polyketide DIF-1.

Song Y, Luciani MF, Giusti C, Golstein P - Mol. Biol. Cell (2014)

Bottom Line: In these cases, c-di-GMP-induced cell death was rescued by complementation with exogenous DIF-1.Taken together, these results demonstrated that c-di-GMP could trigger cell death in Dictyostelium only in the presence of the DIF-1 polyketide or its metabolites.This identified another element of control to this cell death and perhaps also to c-di-GMP effects in other situations and organisms.

View Article: PubMed Central - PubMed

Affiliation: Centre d'Immunologie de Marseille-Luminy, UM2 Aix-Marseille Université; Institut National de la Santé et de la Recherche Médicale, U1104; and Centre National de la Recherche Scientifique, UMR7280, 13288 Marseille, France.

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In contrast to DH1 cells, Dictyostelium HMX44A cells could be induced to die by DIF-1 but not by c-di-GMP. In separate experiments, DH1- or HMX44A-starved cells were subjected to either 100 nM DIF-1 or 10 μM c-di-GMP (A) for 40 h and then photographed (see Figure 1A legend for details) or (B) for 24 h and then subjected to HL5 medium to allow regrowth of surviving cells; regrown cells were counted after 3 more days (see Figure 1B legend for details). (C) HMX44A cells were subjected in LabTek chambers to graded concentrations of DIF-1 and c-di-GMP in a checkerboard manner. The numbers are the percentages of clumps each comprising at least three vacuolized cells after 17 h in the presence or absence of inducers, established as in the legend to Figure 1A. HMX44A cells showed vacuolization when subjected to DIF-1 alone but not to c-di-GMP alone, and there was synergy between the two inducers. These cells made no endogenous DIF-1, leading to no vacuolization by exogenous c-di-GMP. However, addition of exogenous DIF-1 led to vacuolization through the autonomous DIF-1 pathway and through cooperation with exogenous c-di-GMP (see Figure 5 for schematization).
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Figure 2: In contrast to DH1 cells, Dictyostelium HMX44A cells could be induced to die by DIF-1 but not by c-di-GMP. In separate experiments, DH1- or HMX44A-starved cells were subjected to either 100 nM DIF-1 or 10 μM c-di-GMP (A) for 40 h and then photographed (see Figure 1A legend for details) or (B) for 24 h and then subjected to HL5 medium to allow regrowth of surviving cells; regrown cells were counted after 3 more days (see Figure 1B legend for details). (C) HMX44A cells were subjected in LabTek chambers to graded concentrations of DIF-1 and c-di-GMP in a checkerboard manner. The numbers are the percentages of clumps each comprising at least three vacuolized cells after 17 h in the presence or absence of inducers, established as in the legend to Figure 1A. HMX44A cells showed vacuolization when subjected to DIF-1 alone but not to c-di-GMP alone, and there was synergy between the two inducers. These cells made no endogenous DIF-1, leading to no vacuolization by exogenous c-di-GMP. However, addition of exogenous DIF-1 led to vacuolization through the autonomous DIF-1 pathway and through cooperation with exogenous c-di-GMP (see Figure 5 for schematization).

Mentions: We first used Dictyostelium HMX44A cells, which are known to make very little DIF-1 but to remain responsive to exogenous DIF-1 (Kopachik et al., 1983). Accordingly, these cells vacuolized well with exogenous DIF-1 (Figure 2A). In contrast, addition of exogenous c-di-GMP led to small, roundish, contrasted cells with almost no vacuolization (Figure 2A), no cellulose encasings (unpublished data), and no death when tested by regrowth (Figure 2B, right). In addition, in these HMX44A cells, where exogenous c-di-GMP alone did not induce cell death, there was synergy between exogenous DIF-1 and exogenous c-di-GMP (Figure 2C). Taken together, the results indicate that in HMX44A cells relative to DH1 cells, an unknown mutation, or mutations, prevented vacuolar cell death induced by c-di-GMP. Again, irrespective of their other possible defects, HMX44A cells produced no or little DIF-1. These results were in line with the possibility of a requirement of DIF-1 for c-di-GMP induction of cell death.


c-di-GMP induction of Dictyostelium cell death requires the polyketide DIF-1.

Song Y, Luciani MF, Giusti C, Golstein P - Mol. Biol. Cell (2014)

In contrast to DH1 cells, Dictyostelium HMX44A cells could be induced to die by DIF-1 but not by c-di-GMP. In separate experiments, DH1- or HMX44A-starved cells were subjected to either 100 nM DIF-1 or 10 μM c-di-GMP (A) for 40 h and then photographed (see Figure 1A legend for details) or (B) for 24 h and then subjected to HL5 medium to allow regrowth of surviving cells; regrown cells were counted after 3 more days (see Figure 1B legend for details). (C) HMX44A cells were subjected in LabTek chambers to graded concentrations of DIF-1 and c-di-GMP in a checkerboard manner. The numbers are the percentages of clumps each comprising at least three vacuolized cells after 17 h in the presence or absence of inducers, established as in the legend to Figure 1A. HMX44A cells showed vacuolization when subjected to DIF-1 alone but not to c-di-GMP alone, and there was synergy between the two inducers. These cells made no endogenous DIF-1, leading to no vacuolization by exogenous c-di-GMP. However, addition of exogenous DIF-1 led to vacuolization through the autonomous DIF-1 pathway and through cooperation with exogenous c-di-GMP (see Figure 5 for schematization).
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Related In: Results  -  Collection

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Figure 2: In contrast to DH1 cells, Dictyostelium HMX44A cells could be induced to die by DIF-1 but not by c-di-GMP. In separate experiments, DH1- or HMX44A-starved cells were subjected to either 100 nM DIF-1 or 10 μM c-di-GMP (A) for 40 h and then photographed (see Figure 1A legend for details) or (B) for 24 h and then subjected to HL5 medium to allow regrowth of surviving cells; regrown cells were counted after 3 more days (see Figure 1B legend for details). (C) HMX44A cells were subjected in LabTek chambers to graded concentrations of DIF-1 and c-di-GMP in a checkerboard manner. The numbers are the percentages of clumps each comprising at least three vacuolized cells after 17 h in the presence or absence of inducers, established as in the legend to Figure 1A. HMX44A cells showed vacuolization when subjected to DIF-1 alone but not to c-di-GMP alone, and there was synergy between the two inducers. These cells made no endogenous DIF-1, leading to no vacuolization by exogenous c-di-GMP. However, addition of exogenous DIF-1 led to vacuolization through the autonomous DIF-1 pathway and through cooperation with exogenous c-di-GMP (see Figure 5 for schematization).
Mentions: We first used Dictyostelium HMX44A cells, which are known to make very little DIF-1 but to remain responsive to exogenous DIF-1 (Kopachik et al., 1983). Accordingly, these cells vacuolized well with exogenous DIF-1 (Figure 2A). In contrast, addition of exogenous c-di-GMP led to small, roundish, contrasted cells with almost no vacuolization (Figure 2A), no cellulose encasings (unpublished data), and no death when tested by regrowth (Figure 2B, right). In addition, in these HMX44A cells, where exogenous c-di-GMP alone did not induce cell death, there was synergy between exogenous DIF-1 and exogenous c-di-GMP (Figure 2C). Taken together, the results indicate that in HMX44A cells relative to DH1 cells, an unknown mutation, or mutations, prevented vacuolar cell death induced by c-di-GMP. Again, irrespective of their other possible defects, HMX44A cells produced no or little DIF-1. These results were in line with the possibility of a requirement of DIF-1 for c-di-GMP induction of cell death.

Bottom Line: In these cases, c-di-GMP-induced cell death was rescued by complementation with exogenous DIF-1.Taken together, these results demonstrated that c-di-GMP could trigger cell death in Dictyostelium only in the presence of the DIF-1 polyketide or its metabolites.This identified another element of control to this cell death and perhaps also to c-di-GMP effects in other situations and organisms.

View Article: PubMed Central - PubMed

Affiliation: Centre d'Immunologie de Marseille-Luminy, UM2 Aix-Marseille Université; Institut National de la Santé et de la Recherche Médicale, U1104; and Centre National de la Recherche Scientifique, UMR7280, 13288 Marseille, France.

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Related in: MedlinePlus