Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA.

Kuspa, Adam; Loomis, William F.

doi:10.1073/pnas.89.18.8803

Cited by 450 publications

(348 citation statements)

References 25 publications

(19 reference statements)

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“…We took advantage of the powerful genetic tools and the absence of main actors of apoptosis in Dictyostelium to study signaling pathways involved in these cell death types. As reported here, through random insertional mutagenesis (Kuspa and Loomis, 1992) and selection for resistance to death we obtained a Dictyostelium mutant that did not vacuolize and did not undergo ACD. The disrupted gene was iplA, the only gene encoding inositol 1,4,5-trisphosphate (IP 3 ) receptor (IP3R).…”

Section: Introductionmentioning

confidence: 95%

The Inositol 1,4,5-Trisphosphate Receptor Is Required to Signal Autophagic Cell Death

Lam

Kosta

Luciani

et al. 2008

MBoC

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The signaling pathways governing pathophysiologically important autophagic (ACD) and necrotic (NCD) cell death are not entirely known. In the Dictyostelium eukaryote model, which benefits from both unique analytical and genetic advantages and absence of potentially interfering apoptotic machinery, the differentiation factor DIF leads from starvation-induced autophagy to ACD, or, if atg1 is inactivated, to NCD. Here, through random insertional mutagenesis, we found that inactivation of the iplA gene, the only gene encoding an inositol 1,4,5-trisphosphate receptor (IP3R) in this organism, prevented ACD. The IP3R is a ligand-gated channel governing Ca 2؉ efflux from endoplasmic reticulum stores to the cytosol. Accordingly, Ca 2؉ -related drugs also affected DIF signaling leading to ACD. Thus, in this system, a main pathway signaling ACD requires IP3R and further Ca 2؉ -dependent steps. This is one of the first insights in the molecular understanding of a signaling pathway leading to autophagic cell death.

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Section: Introductionmentioning

confidence: 95%

The Inositol 1,4,5-Trisphosphate Receptor Is Required to Signal Autophagic Cell Death

Lam

Kosta

Luciani

et al. 2008

MBoC

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“…Kessin and colleagues (54) did just such a study. They made a large random collection of clones that each had a single gene disrupted by restriction enzyme-mediated integration (REMI), a process that randomly inserts a known sequence containing both restriction cut sites, and an antibiotic resistance gene (55). Kessin and colleagues (54) put a pool of REMI knockouts through 20 generations of selection in a wellmixed (low-relatedness) environment.…”

Section: Dictyostelium Discoideum As a Model System For Cooperationmentioning

confidence: 99%

Evolution of cooperation and control of cheating in a social microbe

Strassmann

Queller

2011

Proc. Natl. Acad. Sci. U.S.A.

194

200

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Much of what we know about the evolution of altruism comes from animals. Here, we show that studying a microbe has yielded unique insights, particularly in understanding how social cheaters are controlled. The social stage of Dictylostelium discoideum occurs when the amoebae run out of their bacterial prey and aggregate into a multicellular, motile slug. This slug forms a fruiting body in which about a fifth of cells die to form a stalk that supports the remaining cells as they form hardy dispersal-ready spores. Because this social stage forms from aggregation, it is analogous to a social group, or a chimeric multicellular organism, and is vulnerable to internal conflict. Advances in cell labeling, microscopy, single-gene knockouts, and genomics, as well as the results of decades of study of D. discoideum as a model for development, allow us to explore the genetic basis of social contests and control of cheaters in unprecedented detail. Cheaters are limited from exploiting other clones by high relatedness, kin discrimination, pleiotropy, noble resistance, and lottery-like role assignment. The active nature of these limits is reflected in the elevated rates of change in social genes compared with nonsocial genes. Despite control of cheaters, some conflict is still expressed in chimeras, with slower movement of slugs, slightly decreased investment in stalk compared with spore cells, and differential contributions to stalk and spores. D. discoideum is rapidly becoming a model system of choice for molecular studies of social evolution.kin selection | kin recognition | multicellularity | major transitions

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“…Isolation of amiB À mutants and structure of the amiB gene To isolate the genes that regulate early developmental stages including the growth/differentiation transition and the aggregation stage in Dictyostelium, we used the restriction enzyme-mediated integration (REMI) (Kuspa & Loomis 1992;Adachi et al 1994) of a blasticidin resistance marker to randomly mutagenize wild-type cells, and isolated a set of aggregate-less mutants. We screened approximately 40 000 REMImutagenized clones for aggregate-less mutant strains that formed smooth plaques on a bacterial lawn, obtaining 28 clones.…”

Section: Resultsmentioning

confidence: 99%

“…Among them, two independent REMI mutants, R8-2 and R12-3, were selected for further analysis in this study because they had insertions in the same gene and displayed severe defects in the aggregation process such as cAMP-induced production of cAMP and chemotaxis to cAMP, as shown below. From R8-2 and R12-3, the marker DNA and¯anking genomic DNA sequences were cloned by the plasmidrescue method (Kuspa & Loomis 1992;Adachi et al 1994) (Fig. 1A).…”

Section: Resultsmentioning

confidence: 99%

amiB, a novel gene required for the growth/differentiation transition in Dictyostelium

2000

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Background: The differentiation programme of Dictyostelium discoideum is initiated by starvation. Nutrient depletion triggers the differentiation of Dictyostelium cells through the transcriptional inactivation of some growth-phase genes, as well as through the transcriptional activation of essential genes required for the aggregation of the cells. The adenylyl cyclase (ACA) gene, acaA, is one of the earliest genes expressed following starvation. ACA produces intracellular and extracellular cAMP that drives further differentiation by inducing chemotaxis, developmental gene expression and morphogenesis of Dictyostelium cells. Although several genes have been identi®ed as being essential for the initiation of differentiation process, such as the transcriptional activation of ACA expression, the molecular mechanisms of the growth/differentiation transition remain to be explored.

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Tagging developmental genes in Dictyostelium by restriction enzyme-mediated integration of plasmid DNA.

Cited by 450 publications

References 25 publications

The Inositol 1,4,5-Trisphosphate Receptor Is Required to Signal Autophagic Cell Death

The Inositol 1,4,5-Trisphosphate Receptor Is Required to Signal Autophagic Cell Death

Evolution of cooperation and control of cheating in a social microbe

amiB, a novel gene required for the growth/differentiation transition in Dictyostelium

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