Hypersensitive sites in the 5′ and 3′ flanking regions of the cysteine proteinase I gene of<i>Dictyostelium discoideum</i>

Pavlovic, Jovan; Banz, Elizabeth; Parish, Roger W.

doi:10.1093/nar/14.22.8703

Cited by 14 publications

(16 citation statements)

References 43 publications

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“…While this paper was under revision, Pavlovic et al reported the identification of hypersensitive sites in the 5' and 3' flanking regions of the D. discoideum cysteine proteinase I gene (25). Similar to the situation reported here, the hypersensitive sites associated with the 5' end of this gene are present only when the gene is activated.…”

Section: Resultssupporting

confidence: 76%

Developmental regulation of DNase I-hypersensitive sites in Dictyostelium discoideum.

Ayres¹,

Neuman²,

Rowekamp³

et al. 1987

Mol. Cell. Biol.

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We have studied two regions of Dictyostelium discoideum chromatin and identified several DNase I-hypersensitive sites in these regions. One of these sites is located about 300 to 500 bases upstream of the transcriptional start site of a gene that is expressed at all stages of development. This site is present in both vegetative cells and postaggregation cells. Another hypersensitive site is associated with a gene that is expressed only after the multicellular stage. This site is located about 400 bases upstream of the start site, and it is present only in postaggregation cells. Thus, much like higher eucaryotes, D. discoideum contains DNase I-hypersensitive sites that may be involved in the regulation of the genes with which they are associated.

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Section: Resultssupporting

confidence: 76%

Developmental regulation of DNase I-hypersensitive sites in Dictyostelium discoideum.

Ayres¹,

Neuman²,

Rowekamp³

et al. 1987

Mol. Cell. Biol.

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“…It seems likely that the sequences required for the expression of the DG17 and CP1 genes lie within the G-rich regions, given their unusual sequence, position, and hypersensitivity to nucleases during development (32). Short G+C rich regions are found upstream of a number of other cAMP-inducible genes in Dictyostelium spp.…”

Section: Discussionmentioning

confidence: 99%

“…More direct evidence for this came from studies on the structure of the chromatin flanking the CP1 gene. Four sites have been identified upstream of the CP1 gene at -220, -300, -670, and -770 relative to the start site of transcription, which became hypersensitive to nucleases during aggregation, when the CP1 gene is transcribed (32). When we compared the positions of these hypersensitive sites with the sequence of the intergenic region, these sites mapped to the four G-rich boxes (Fig.…”

Section: Methodsmentioning

confidence: 99%

Two divergently transcribed genes of Dictyostelium discoideum are cyclic AMP-inducible and coregulated during development.

Driscoll¹,

Williams²

1987

Mol. Cell. Biol.

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The cysteine proteinase 1 (CP1) gene of Dictyostelium discoideum encodes a developmentally regulated sulfhydryl proteinase. We characterized the DNA sequences upstream of the CP1 gene and found a second developmentally regulated gene, which we term DG17. The translational open reading frame of the DG17 gene encoded a 458-amino-acid cysteine-and lysine-rich protein of unknown function. In several regions, the cysteine and lysine residues were arranged in a manner characteristic of the zinc-binding domains found in proteins which interact with nucleic acids. During normal development, the DG17 and CP1 genes are coordinately activated late in aggregation. The addition of exogenous cyclic AMP (cAMP) induced the premature expression of both mRNAs. By measuring the rate of specific mRNA synthesis in isolated nuclei, we showed that cAMP acted at the transcriptional level to activate both genes. The two genes were separated by 910 nucleotides and were divergently transcribed. The intergenic region was predominantly composed of A+T residues except for four short G-rich regions. These sequences coincided with the positions of four nuclease-hypersensitive sites, which appear during aggregation when the DG17 and CP1 genes are transcribed (J. Pavlovic, E. Banz, and R. W. Parish, Nucleic Acids Res. 14: [8703][8704][8705][8706][8707][8708][8709][8710][8711][8712][8713][8714][8715][8716][8717][8718][8719][8720][8721][8722] 1986). Two of the G-rich regions formed the core of two almost identical 80-nucleotide repeats located 220 and 320 nucleotides upstream of the CP1 gene. Using the Dictyostelium transformation system, we showed that a restriction fragment containing the intergenic region was capable of directing bidirectional transcription in a cAMP-dependent manner.In response to starvation, single cells of Dictyostelium discoideum form aggregates which develop into multicellular fruiting bodies consisting of stalk and spore cells. Aggregation occurs in response to pulsatile emissions of cyclic AMP (cAMP) from a signalling center (22). As multicellular aggregates form, there are major changes in gene expression to yield the new proteins required for cellular differentiation, and approximately 2,000 to 3,000 mRNAs are induced (1,4,5). In addition to its role as a chemoattractant, the rise in extracellular cAMP concentration during aggregation also plays a role in regulating gene expression. The addition of exogenous cAMP to developing cells represses the expression of a number of genes active early in development and induces the premature expression of a group of genes which are active late during aggregation (13,21,26,28,40,46). Although it has been shown that cAMP can alter the rate of transcription of specific genes in D. discoideum (21, 46), the precise mechanism of action of cAMP is not understood. Pharmacological studies suggest that the effects of extracellular cAMP on gene expression are mediated by the cell surface cAMP receptor (12,24,31). In the postaggregative stage, cAMP plays an additional role in controlling gen...

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“…Cold Spring Harbor Laboratory Press on May 10, 2018 -Published by genesdev.cshlp.org Downloaded from gene is first expressed (Pavlovic et al 1986;Driscoll and Williams 1987). Of these two elements, the gene-proximal one (B4) shows a striking resemblance to an inverted copy of the CP1 G-rich element, whereas the DG17-B3 element bears no apparent sequence homology to either the pst-cath/CP2 or the CP1 G-rich elements.…”

Section: Genes and Development 421mentioning

confidence: 99%

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

Abstract: The cysteine proteinase

Cited by 14 publications

References 43 publications

Developmental regulation of DNase I-hypersensitive sites in Dictyostelium discoideum.

Developmental regulation of DNase I-hypersensitive sites in Dictyostelium discoideum.

Two divergently transcribed genes of Dictyostelium discoideum are cyclic AMP-inducible and coregulated during development.

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

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