A developmentally regulated cysteine proteinase in Dictyostelium discoideum.

Williams, Jeffrey; North, Michael; Mahbubani, Hiro

doi:10.1002/j.1460-2075.1985.tb03730.x

Cited by 71 publications

(46 citation statements)

References 43 publications

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“…Although all of these genes have been shown to be transcriptionally induced by cAMP and show similar developmental kinetics of expression, the only apparent similarity between the putative regulatory elements is that they are G/C rich (Williams et al 1980;Mehdy et al 1983;Mehdy and Firtel 1985;Pears et al 1985;Williams et al 1985;Datta et al 1986;Driscoll and Williams 1987;Pavlovic et al 1989). The rest of the 5'-flanking regions are >90% A/T, similar to the majority of the 5'-flanking regions of other Dictyostelium genes (Kimmel and Firtel 1983).…”

Section: Structure Of the G/c-rich Elementsmentioning

confidence: 94%

A developmentally regulated trans-acting factor recognizes dissimilar G/C-rich elements controlling a class of cAMP-inducible Dictyostelium genes.

Hjorth¹,

Pears²,

Williams³

et al. 1990

Genes Dev.

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Transcriptional response elements involved in the cAMP-inducible and developmentally regulated expression of the Dictyostelium aggregate-stage gene pst-cath/CP2 have been shown to include a G/C-rich sequence element [G-box regulatory element {GBRE1]. We have recently identified a trans-acting factor, GBF (GBRE binding factor), that specifically interacts with this sequence and have shown that the binding activity of GBF to GBRE is developmentally regulated and inducible by cAMP. Here, we examine further the possible role of GBF in the regulation of pst-cath/CP2 and three other coordinately regulated, cAMP-inducible aggregate-stage genes. We show that GBF itself {or other closely related factorsl recognizes dissimilar G/C-rich elements present in the 5'-flanking regions of these genes and that the ability of the individual, distinct G/C-rich elements to confer regulated expression on a promoter deletion mutant of the pst-cath/CP2 gene is correlated with the relative affinity for GBF. G/C-rich elements carrying point mutations that prevent in vitro binding of GBF to two of the G/C-rich elements fail to activate expression in vivo. An analysis of major points of contact between the GBF protein and two distinctly different binding sites suggests that binding of GBF to these sequence elements involves a considerable degree of flexibility in DNA-protein interactions. These results suggest that the regulated expression of a class of aggregate-stage cAMP-inducible genes involves the interaction of GBF or homologous factors with dissimilar G/C-rich sequence elements and that induction of GBF activity or that of homologous factors by cAMP may thus be a limiting step in the induction of this temporally coordinate set of genes during Dictyostelium development. Dictyostelium discoideum propagates as a vegetative amoeba in the presence of nutrients and initiates, upon starvation, a multicellular differentiation process that results in the formation of a mature fruiting body containing two distinctly different cell types--stalk cells and spores. Within several hours of starvation, Dictyostelium cells aggregate in response to pulsatile cAMP signals. Binding of cAMP to cell-surface receptors activates adenylate cyclase, and repeated cycles of cAMP synthesis and secretion serve to transmit, or relay, the chemotactic signal outward through the population of aggregating cells (see Janssens and van Haastert 1987;Firtel et al. 1989). After formation of tightly associated multicellular aggregates, a migrating pseudoplasmodium (slug) is formed in which precursor cells to the 3Corresponding author.

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Section: Structure Of the G/c-rich Elementsmentioning

confidence: 94%

A developmentally regulated trans-acting factor recognizes dissimilar G/C-rich elements controlling a class of cAMP-inducible Dictyostelium genes.

Hjorth¹,

Pears²,

Williams³

et al. 1990

Genes Dev.

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“…The net cysteine proteinase activity is greatest during the vegetative phase, but declines during development as aggregation and then fruiting body formation take place. Cysteine proteinase genes which are not expressed during growth are switched on during this period (Pears et al, 1985;Williams et al, 1985); however, the products of these genes have not yet been identified. Spores have very low cysteine proteinase activity, but activity accumulates during germination (Jackson & Cotter, 1984;North & Cotter, 1984).…”

Section: Introductionmentioning

confidence: 99%

Dictyostelium discoideum spore germination: increases in proteinase activity are not directly coupled to the emergence of myxamoebae

North

Cotter

Franek

1990

Journal of General Microbiology

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The accumulation of proteinase activity during the germination of Dictyostelium discoideum spores was examined under conditions in which the timing of germination events was varied. Spores had a single major proteinase, the aspartic proteinase ddAP58 (proteinase E), while the extracellular matrix which surrounds the spores in the fruiting body contained a number of lower-Mr cysteine proteinases, the most active being ddCP18. Very little peptide-nitroanilide-hydrolysing activity was detectable in spores, although some was associated with the matrix. During spore germination there were large increases in activity towards N-carbobenzoxy-L-tyrosyl-L-lysyl-Larginine 4-nitroanilide and N-benzoyl-L-prolyl-L-phenylalanyl-L-arginine 4-nitroanilide. The enzymes responsible for these activities (referred to as ZYKRase and BzPFRase respectively) were not identical as BzPFRase was much more sensitive to the cysteine proteinase inhibitor E-64 than was ZYKRase. When spores were heatactivated, the increases in activity coincided with the emergence of myxamoebae and the appearance of cysteine proteinases detected using electrophoresis in gelatin gels. For autoactivated spores, emergence was delayed by 0.5 to 1 h and proteinase accumulation lagged slightly behind emergence. If the spores were activated with DMSO, or if heat-activated spores were treated with sucrose after swelling, proteinase accumulation proceeded more rapidly than emergence. Thus temporal control of the accumulation of proteinases during germination varies according to the conditions used. In heat-activated spores, the timing of the increase was similar to that observed previously for P-glucosidase and trehalase, but the temporal controls were not the same as those for the latter enzymes when the other activation conditions were used. The results show that proteinase accumulation is not directly coupled to emergence but could be more closely linked to the late swelling stage which precedes emergence.

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“…The gene is developmentally regulated, transcription commencing at the 6th hour of development and ceasing after 12 h of differentiation (Pavlovic et al submitted). Messenger RNA accumulation reaches a maximum between 8 and 12 h when it constitutes about 1% of the mRNA population (32,33). The messenger RNA can be accumulated prematurely when exogenous cAMP is present (34).…”

Section: Introductionmentioning

confidence: 99%

“…The messenger RNA can be accumulated prematurely when exogenous cAMP is present (34). It is approximately 1200 nucleotides in length and encodes a 36,000 dalton polypeptide (cysteine proteinase I) with a high degree of homology to plant and animal cysteine proteinases (33). Using the indirect end-labeling technique (2,35) we have mapped the micrococcal nuclease, DNase I and endogenous hypersensitive sites at the 5' and 3' flanking regions of the gene.…”

Section: Introductionmentioning

confidence: 99%

Hypersensitive sites in the 5′ and 3′ flanking regions of the cysteine proteinase I gene ofDictyostelium discoideum

1986

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A developmentally regulated cysteine proteinase in Dictyostelium discoideum.

Cited by 71 publications

References 43 publications

A developmentally regulated trans-acting factor recognizes dissimilar G/C-rich elements controlling a class of cAMP-inducible Dictyostelium genes.

A developmentally regulated trans-acting factor recognizes dissimilar G/C-rich elements controlling a class of cAMP-inducible Dictyostelium genes.

Dictyostelium discoideum spore germination: increases in proteinase activity are not directly coupled to the emergence of myxamoebae

Hypersensitive sites in the 5′ and 3′ flanking regions of the cysteine proteinase I gene ofDictyostelium discoideum

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