Induction and repression of the Drosophila Sgs-3 glue gene are mediated by distinct sequences in the proximal promoter.

Martin, Marianne; Giangrande, Angela; Ruiz, Claude; Richards, Gareth

doi:10.1002/j.1460-2075.1989.tb03410.x

Cited by 31 publications

(16 citation statements)

References 37 publications

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“…Since there might also be heterologous interactions with other site 2 proteins bound farther upstream, one might expect a fairly complex array of levels of expression, depending on how many binding sites for each of the two factors is present in a given locus. This is in fact what has been seen by us and others (28,43): in constructs where upstream binding sites are still present, deleting part or all of sites 1 or 2 leaves varying amounts of residual activity. In this context it is interesting to note the point mutant PM(-90).…”

Section: Discussionsupporting

confidence: 79%

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Fine-structure mutational analysis of a stage- and tissue-specific promoter element of the Drosophila glue gene Sgs-3.

Todo¹,

Roark²,

Raghavan³

et al. 1990

Mol. Cell. Biol.

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The Sgs-3 gene of Drosophila melanogaster exhibits a tightly regulated pattern of expression governed by two functionally equivalent elements within 1 kb of the gene, each of which is sufficient to confer third-instar salivary gland-specific transcription. In this report we describe a detailed functional analysis of one of these, the proximal element. To determine the nucleotides responsible for specific expression, we have introduced mutations into the proximal element and then assessed the effects of each alteration on expression in the developing animal. We have identified six particularly important base pairs which are located in two regions separated by nonessential sequences. These base pairs, along with some surrounding sequence, are conserved within the upstream regions of the three glue genes at 68C. Nearly identical groups of base pairs can be found upstream of the other glue genes which have been cloned. This analysis has allowed us to derive a consensus sequence, which we believe contains binding sites for two different factors which interact to direct third-instar salivary gland-specific expression.The Sgs-3 gene of Drosophila melanogaster encodes a glycoprotein component of the secretion produced in the salivary glands of the third-instar larva, which is expelled just prior to pupariation (1, 18). This "glue" serves to affix the pupa to a surface for the duration of metamorphosis (8). The Sgs-3 gene is one of three glue genes clustered at 68C on the left arm of the third chromosome (9, 32). Two other of the eight known glue genes, Sgs4 at 3C and Sgs-5 at 90BC, have also been cloned (34). The Sgs-3 gene exhibits a tightly regulated pattern of expression. It is abundantly transcribed only in a single cell type, of a single tissue, during a single 30-h period of development: the secretory cells of the salivary gland in the mid-to late third larval instar. Our interest has been in understanding how this extraordinarily specific expression pattern is achieved.Two trans-acting regulators of this gene have been previously described (see reference 33 for a review). The steroid hormone ecdysterone must be present at the time of the second molt, or no Sgs-3 mRNA is produced (14). In addition, particular mutations at the 2B5 locus, such as that of the nonpupariating mutant npr-13, fail to produce Sgs-3 mRNA (5). Whether either of these factors acts directly at the Sgs-3 locus is not yet known.The required cis-acting sequences have been investigated in our laboratory and by others (for reviews, see references 29 and 33). An element located within 130 bp of the transcription start site (the proximal element) is sufficient for correctly regulated but low-level expression of this gene (54). A second element, located between -130 and -629 bp relative to the transcription start site (the distal element), is also sufficient for regulated low-level expression (43). When these elements are both present, a 20-fold enhancement of the level of expression is seen. Within the proximal element,

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Section: Discussionsupporting

confidence: 79%

“…When these elements are both present, a 20-fold enhancement of the level of expression is seen. Within the proximal element, only sequences between -56 and -98 are required for full function (28,43).…”

mentioning

confidence: 99%

Fine-structure mutational analysis of a stage- and tissue-specific promoter element of the Drosophila glue gene Sgs-3.

Todo¹,

Roark²,

Raghavan³

et al. 1990

Mol. Cell. Biol.

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“…The hsp70 proximal promoter (from the TATA box} is compatible with upstream elements and enhancers of other genes. The fat body enhancer element of the Drosophila YP1 gene is effective on the hsp70 proximal promoter (Garabedian et al 1986), as are the elements of the Drosophila Sgs-3 gene (Martin et al 1989). Likewise, a model transcriptional inducer from yeast, GAL4, when expressed in Drosophila, will induce an hsp70-derived promoter that has nearby DNA regulatory sequences that bind GAL4 (Fischer et al 1988).…”

Section: Perspectives On Polymerase Pausingmentioning

confidence: 99%

Promoter melting and TFIID complexes on Drosophila genes in vivo.

Giardina¹,

Pérez‐Riba²,

Lis³

1992

Genes Dev.

146

162

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In vivo UV cross-linking and nuclear transcriptional run-on experiments have shown that a number ofDrosophila genes possess an elongationally paused RNA polymerase on their 5' ends. Here, we examine in vivo promoters that do and do not possess paused polymerases using the single-stranded DNA-probing reagent KMnO 4. Melted DNA helices are found associated with the pause site of the uninduced hspTO and hsp26 heat shock genes and the constitutively expressed I~-1 tubulin gene. The histone 1-11 and H2B genes, which lack a paused polymerase, have no comparable region of melted DNA. Melting at the pause site persists upon heat shock induction of the hsp 70 and hsp26 genes, indicating that pausing continues after gene activation.Interestingly, activation triggers additional melting, both at the start site (in the region where open complexes would be expected to form) and downstream of the uninduced pause site. In the course of our studies, we discovered that some T residues of the TATA box were protected from KMnO4 modification in both induced and uninduced cells. This protection appears to be a consequence of TFIID binding, as a similar protection pattern could be produced in vitro with purified protein.

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“…Many of these regulatory proteins possess similar structural domains that mediate binding to specific DNA sequences (9,21,22,32,33) and are, in at least some cases, encoded by members of large evolutionarily related gene families (1,22 shock-induced developmental expression of the introduced hsp7O-lacZ genes that are conferred by the synthetic regulatory sequences. We show that each of the three synthetic sequence variants of the heat shock element that we examined confers a distinct pattern of developmental expression in transgenic Drosophila lines.The heat shock gene hsp7O in D. melanogaster appears to be exclusively under heat shock control (16, 27, 41); however, the hsp7O promoter that is deleted of its native heat shock regulatory sequence has been shown to be a very receptive target for activation by a variety of regulatory elements (7,10,12,26). Figure 1A shows the structure of the hsp7O-lacZ transposons used in this study.…”

mentioning

confidence: 99%

“…The heat shock gene hsp7O in D. melanogaster appears to be exclusively under heat shock control (16,27,41); however, the hsp7O promoter that is deleted of its native heat shock regulatory sequence has been shown to be a very receptive target for activation by a variety of regulatory elements (7,10,12,26). Figure 1A shows the structure of the hsp7O-lacZ transposons used in this study.…”

mentioning

confidence: 99%

Closely related DNA sequences specify distinct patterns of developmental expression in Drosophila melanogaster.

Xiao

Lis

1990

Mol. Cell. Biol.

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Three short synthetic DNA sequences, which are closely related to one another, confer three distinct patterns of developmental expression on the heat shock hsp7O gene in transgenic Drosophila melanogaster lines. These results show that small variations or even single base pair changes in a repeated element of a regulatory sequence can create promoters that display new specificities of tissue and developmental regulation. Interestingly, the three patterns of developmental expression conferred by the synthetic DNAs resemble in part those of the known developmental genes: glucose dehydrogenase (Gld), Dopa decarboxylase (Ddc), and salivary gland secretory proteins (Sgs), respectively. In each case, the defined regulatory region of the known developmental gene contains multiple sequences that are similar or identical to the synthetic sequence that confers a similar pattern of developmental expression on the hsp7O gene. Thus, these results are congruent with the view that short sequence elements in multiple copies can confer either simple or relatively complex patterns of developmental expression on a receptive promoter like that of hsp7O. Furthermore, the fact that the three variants tested produced three distinct patterns of expression in transgenic animals suggests that the number of different elements is large.Regulatory regions of genes contain multiple short sequence elements that are binding sites for specific regulatory proteins (19,24,32). These binding interactions are determined in large part by both the affinity of the proteins for particular sequence elements and the abundance of these proteins in the particular cell type in question. The effect of these protein-DNA interactions on transcription depends on the transcriptional activation (or repression) strength of a protein and on the constellation of such protein-DNA complexes that are associated with a particular gene (32). The number of distinct regulatory proteins that have been identified to interact with specific DNA sequence elements is large and growing rapidly, particularly in the cases of multicellular organisms (1,9,15,19,21,22,24,32,35). Many of these regulatory proteins possess similar structural domains that mediate binding to specific DNA sequences (9,21,22,32,33) and are, in at least some cases, encoded by members of large evolutionarily related gene families (1,22 shock-induced developmental expression of the introduced hsp7O-lacZ genes that are conferred by the synthetic regulatory sequences. We show that each of the three synthetic sequence variants of the heat shock element that we examined confers a distinct pattern of developmental expression in transgenic Drosophila lines.The heat shock gene hsp7O in D. melanogaster appears to be exclusively under heat shock control (16, 27, 41); however, the hsp7O promoter that is deleted of its native heat shock regulatory sequence has been shown to be a very receptive target for activation by a variety of regulatory elements (7,10,12,26). Figure 1A shows the structure of the hsp7O-lacZ transposon...

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Induction and repression of the Drosophila Sgs-3 glue gene are mediated by distinct sequences in the proximal promoter.

Cited by 31 publications

References 37 publications

Fine-structure mutational analysis of a stage- and tissue-specific promoter element of the Drosophila glue gene Sgs-3.

Fine-structure mutational analysis of a stage- and tissue-specific promoter element of the Drosophila glue gene Sgs-3.

Promoter melting and TFIID complexes on Drosophila genes in vivo.

Closely related DNA sequences specify distinct patterns of developmental expression in Drosophila melanogaster.

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