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A steroid-triggered transcriptional hierarchy controls salivary gland cell death during Drosophila metamorphosis

MOLECULAR CELL 2000年第3期5卷 445-455页

作者： Jiang, CA Lamblin, AFJ Steller, H Thummel, CS Univ Utah Dept Human Genet Howard Hughes Med Inst Salt Lake City UT 84112 USA MIT Dept Biol Howard Hughes Med Inst Cambridge MA 02139 USA

The steroid hormone ecdysone signals the stage-specific programmed cell death of the larval salivary glands during Drosophila metamorphosis. This response is preceded by an ecdysone-triggered switch in gene expression in which the diap2 death inhibitor is repressed and the reaper (rpr) and head involution defective (hid) death activators are induced. Here we show that rpr is induced directly by the ecdysonereceptor complex through an essential response element in the rpr promoter. The Broad-Complex (BR-C) is required for both rpr and hid transcription, while E74A is required for maximal levels of hid induction. diap2 induction is dependent on beta FTZ-F1, while E75A and E75B are each sufficient to repress diap2. This study identifies transcriptional regulators of programmed cell death in Drosophila and provides a direct link between a steroid signal and a programmed cell death response.

关键词：细胞凋亡/遗传学 DNA结合蛋白质类/代谢二聚作用果蝇属/胚胎学果蝇属/遗传学果蝇蛋白质类蜕皮激素/药理学 fushitarazu转录因子类基因表达调控发育期同源盒结构域蛋白质类凋亡抑制蛋白质类昆虫蛋白质类/遗传学昆虫蛋白质类/代谢变态生物学神经肽类/遗传学肽类/遗传学受体胞质和核受体类固醇/代谢反应元件涎腺/细胞学信号传导类固醇生成因子1 转录因子/代谢转录遗传动物

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引用

CHROMOSOMA 1976年第2期58卷 137-153页

作者： PANKOW, W LEZZI, M HOLDEREGGERMAHLING, I ETH INST ZELL BIOL CH-8093 ZURICH SWITZERLAND

Salivary glands of various stages of the last larval instar of C. tentans were quantitatively analyzed with respect to the expansion of their Balbiani rings (B1, B2, B3) by a fast green staining procedure as well as to the rate of synthesis of their secretory proteins (S1, S2, S3) by a scintillation counting procedure of electrophoretic fractions. The extent of expansion of B1, B2 and B3 correlates positively with the rate of synthesis of S3, S2 and S1, respectively. With B1 and S3 these parameters undergo a parallel and developmentally specific change being rather depressed in intermolt, and particularly in diapausing animals.

关键词：染色体双翅目/代谢蜕皮激素/药理学涎腺/代谢涎腺/超微结构唾液蛋白质类和肽类/生物合成动物

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BETA-GALACTOSIDASE IS INDUCED BY HORMONE IN DROSOPHILA-MELANOGASTER CELL-CULTURES

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 1978年第12期75卷 6102-6106页

作者： BESTBELPOMME, M COURGEON, AM RAMBACH, A INST PASTEUR INSERM UNITE GENIE GENET F-75015 PARIS FRANCE

D. melanogaster cell lines Kc and Ca and clones FC and RF6, cultured in vitro, have no detectable .beta.-galactosidase (.beta.-D-galactoside galactohydrolase, EC 3.2.1.23) activity (as measured by hydrolysis of o-nitrophenyl-.beta.-D-galactoside). Ecdysterone clearly induces .beta.-galactosidase activity in D. melanogaster cells cultured in vitro. Induction occurs in cell lines or clones sensitive to ecdysterone (K, Ca, and Fc) and does not occur in clones resistant to the hormone (RF6). Some properties of the hormone-induced .beta.-galactosidase activity were studied. The Km for o-nitrophenyl galactoside is 0.35 mM and the Ki for lactose is 12 mM (similar to those of Escherichia coli .beta.-galactosidase);the activity can be recovered after sodium dodecyl sulfate treatment;the enzyme is a tetramer (Mr [molecular ratio] of the monomer is 64,000).

关键词：剂量效应关系药物黑腹果蝇/酶学蜕皮激素/药理学蜕皮甾酮/药理学酶诱导/药物作用半乳糖苷酶类/生物合成动力学分子量构效关系 β半乳糖苷酶类/生物合成 β半乳糖苷酶类/遗传学

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Application of exogenously regulatable promoter systems to transgenic models for the study of aging

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JOURNALS OF GERONTOLOGY SERIES A-BIOLOGICAL SCIENCES AND MEDICAL SCIENCES 1999年第1期54卷 B30-B40页

作者： Morgan, WW Richardson, A Sharp, ZD Walter, CA Univ Texas Hlth Sci Ctr Dept Cellular & Struct Biol San Antonio TX 78284 USA Univ Texas Hlth Sci Ctr Dept Physiol San Antonio TX 78284 USA Univ Texas Hlth Sci Ctr Dept Mol Med San Antonio TX 78284 USA Univ Texas Hlth Sci Ctr Inst Biotechnol San Antonio TX 78284 USA S Texas Vet Hlth Care Syst Ctr Geriatr Res Educ & Clin San Antonio TX USA

Transgenic mouse and gene knockout technologies offer powerful tools for dissecting the roles of specific genes in the process of aging. The interpretation of the results of such studies is limited, however by the fact that the gene of interest is over- or underexpressed throughout the life span of the animal model. Among other problems, this situation makes it difficult to separate the effects that a specific gene has on embryological development from those that it may exert on Me subsequent maturation and aging of the animal. It is also not possible with these methods alone to alter the expression of genes ill an age-dependent fashion and to assess the effects of these alterations on the aging process. This capacity would be of particular interest in studying genes which are thought to have a role in regulating physiological homeostasis. Because they offer the opportunity to activate or render inactive the expression of genes at will, exogenously regulatable promoter systems, particularly when used in combination with traditional transgenic or gene knockout approaches, provide a new and potentially very powerful tool for studying the effect of selected genes on aging. This review discusses the merits and limitations of Be application of either the tetracycline-regulatable promoter system, the RU486-inducible promoter system, or the ecdysone-inducible promoter system to exogenously regulate the expression of a transcriptionally linked gene and to thus assess the effect of that gene on aging.

关键词：衰老/遗传学疾病模型动物蜕皮激素/药理学胚胎和胎儿发育/遗传学基因表达调控/药物作用基因表达调控/遗传学生长/遗传学内环境稳定/遗传学小鼠基因敲除/遗传学小鼠转基因/遗传学米非司酮/药理学启动区(遗传学)/药物作用启动区(遗传学)/遗传学四环素/药理学反式激活(遗传学)/药物作用反式激活(遗传学)/遗传学转录遗传/药物作用转录遗传/遗传学动物小鼠

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Dual requirement for the EcR USP nuclear receptor and the dGATAb factor in an ecdysone response in Drosophila melanogaster

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MOLECULAR AND CELLULAR BIOLOGY 1999年第8期19卷 5732-5742页

作者： Brodu, V Mugat, B Roignant, JY Lepesant, JA Antoniewski, C Univ Paris 06 Inst Jacques Monod Dev Biol Lab CNRSUMR 7592 F-75251 Paris 05 France Univ Paris 07 Inst Jacques Monod Dev Biol Lab CNRSUMR 7592 F-75251 Paris France

The EcR/USP nuclear receptor controls Drosophila metamorphosis by activating complex cascades of gene transcription in response to pulses of the steroid hormone ecdysone at the end of larval development. Ecdysone release provides a ubiquitous signal for the activation of the receptor, but a number of its target genes are induced in a tissue- and stage-specific manner. Little is known about the molecular mechanisms involved in this developmental modulation of the EcR/USP-mediated pathway. Fbp1 is a good model of primary ecdysone response gene expressed in the fat body for addressing this question. We show here that the dGATAb factor binds to three target sites flanking an EcR/USP binding site in a 70-bp enhancer that controls the tissue and stage specificity of Fbp1 transcription. We demonstrate that one of these sites and proper expression of dGATAb are required for specific activation of the enhancer in the fat body. In addition, we provide further evidence that EcR/USP plays an essential role as a hormonal timer. Our study provides a striking example of the integration of molecular pathways at the level of a tissue-specific hormone response unit.

关键词：碱基序列结合部位 DNA结合蛋白质类/生理学果蝇蛋白质类黑腹果蝇/遗传学黑腹果蝇/生长和发育蜕皮激素/药理学增强子元件遗传学脂肪体/代谢真菌蛋白质类/生理学 GATA转录因子类基因表达调控发育期/生理学昆虫蛋白质类/生物合成昆虫蛋白质类/遗传学幼虫大分子物质分子序列数据器官特异性蛋白质结合酿酒酵母蛋白质类时间因素转录因子/生理学动物

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Molecular determinants of differential ligand sensitivities of insect ecdysteroid receptors

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MOLECULAR AND CELLULAR BIOLOGY 2000年第11期20卷 3870-3879页

作者： Wang, SF Ayer, S Segraves, WA Williams, DR Raikhel, AS Michigan State Univ Dept Entomol Genet Program E Lansing MI 48824 USA Michigan State Univ Dept Entomol Cell Biol Program E Lansing MI 48824 USA Michigan State Univ Dept Entomol Program Mol Biol E Lansing MI 48824 USA Yale Univ Dept Mol Cellular & Dev Biol New Haven CT 06520 USA Univ Liverpool Sch Biol Sci Liverpool L69 7ZB Merseyside England

The functional receptor for insect ecdysteroid hormones is a heterodimer consisting of two nuclear hormone receptors, ecdysteroid receptor (EcR) and the retinoid X receptor homologue Ultraspiracle (USP). Although ecdysone is commonly thought to be a hormone precursor and 20-hydroxyecdysone (20E), the physiologically active steroid, little is known about the relative activity of ecdysteroids in various arthropods. As a step toward characterization of potential differential ligand recognition, we have analyzed the activities of various ecdysteroids using gel mobility shift assays and transfection assays in Schneider-2 (S2) cells. Ecdysone showed little activation of the Drosophila melanogaster receptor complex (DmEcR-USP). In contrast, this steroid functioned as a potent ligand for the mosquito Aedes aegypti receptor complex (AaEcR-USP), significantly enhancing DNA binding and transactivating a reporter gene in S2 cells. The mosquito receptor also displayed higher hormone-independent DNA binding activity than the Drosophila receptor. Subunit-swapping experiments indicated that the EcR protein, not the USP protein, was responsible for ligand specificity. Using domain-swapping techniques, we made a series of Aedes and Drosophila EcR chimeric constructs. Differential ligand responsiveness was mapped near the C terminus of the ligand binding domain, within the identity box previously implicated in the dimerization specificity of nuclear receptors. This region includes helices 9 and 10, as determined by comparison with available crystal structures obtained from other nuclear receptors. Site-directed mutagenesis revealed that Phe529 in Aedes EcR, corresponding to Tyr611 in Drosophila EcR, was most critical for ligand specificity and hormone-independent DNA binding activity. These results demonstrated that ecdysone could function as a bona fide ligand in a species-specific manner.

关键词：伊蚊属氨基酸序列细胞系染色体图 DNA/代谢 DNA结合蛋白质类/遗传学 DNA结合蛋白质类/代谢果蝇蛋白质类黑腹果蝇蜕皮激素/代谢蜕皮激素/药理学蜕皮甾酮/代谢蜕皮甾酮/药理学配体分子序列数据苯丙氨酸/遗传学苯丙氨酸/代谢受体类固醇/遗传学受体类固醇/代谢序列同源性氨基酸转录因子/遗传学转录因子/代谢转录激活酪氨酸/遗传学酪氨酸/代谢动物

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BETA-ECDYSONE EFFECTS ON DEVELOPMENT OF NUCLEOPOLYHEDROSIS IN HELIOTHIS SPP

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JOURNAL OF INVERTEBRATE PATHOLOGY 1975年第1期26卷 121-123页

作者： KEELEY, LL VINSON, SB TEXAS A&M UNIV DEPT ENTOMOLAGR EXPT STNCOLLEGE STNTX 77843

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The E23 early gene of Drosophila encodes an ecdysone-inducible ATP-binding cassette transporter capable of repressing ecdysone-mediated gene activation

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PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 2000年第17期97卷 9519-9524页

作者： Hock, T Cottrill, T Keegan, J Garza, D Florida State Univ Dept Sci Biol Tallahassee FL 32306 USA Florida State Univ Med Sci Program Tallahassee FL 32306 USA

At the onset of Drosophila metamorphosis, the steroid hormone 20-OH ecdysone directly induces a small number of early puffs in the polytene chromosomes of the larval salivary gland. Proteins encoded by the early genes corresponding to these transcriptional puffs then regulate the activity of both the early puffs themselves and a much larger set of late puffs. Three of these early genes encode transcription factors that play critical regulatory roles during metamorphosis. Here we report the cloning, DNA sequence, genomic structure, ecdysone inducibility, and temporal expression of an early gene residing in the 23E early puff and denoted E23 (Early gene at 23). In contrast to other early genes, E23 encodes a protein with similarity to ATP-binding cassette transporters. Using heat shock-inducible transgenes, we found that E23 overexpression not only produces phenotypic abnormalities and lethality, but also interferes with ecdysone-mediated gene activation, demonstrating that E23 is capable of modulating the ecdysone response. Our results suggest the existence of a previously unrecognized regulatory mechanism for modulating steroid hormone signaling in Drosophila.

关键词： ATP结合匣式转运子/化学 ATP结合匣式转运子/遗传学 ATP结合匣式转运子/代谢氨基酸序列克隆分子果蝇蛋白质类黑腹果蝇/药物作用黑腹果蝇/遗传学黑腹果蝇/生长和发育黑腹果蝇/代谢蜕皮激素/拮抗剂和抑制剂蜕皮激素/药理学基因表达谱基因昆虫/遗传学基因致死/遗传学昆虫蛋白质类/化学昆虫蛋白质类/遗传学昆虫蛋白质类/代谢幼虫/药物作用幼虫/遗传学变态生物学/药物作用变态生物学/遗传学分子序列数据多基因族/遗传学表型蛹/药物作用蛹/遗传学 RNA 信使/遗传学 RNA 信使/代谢序列比对转录遗传/药物作用转录遗传/遗传学转录激活/药物作用动物

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SEQUENTIAL GENE ACTIVATION BY ECDYSONE IN POLYTENE CHROMOSOMES OF DROSOPHILA-MELANOGASTER .4. INHIBITION BY JUVENILE HORMONES

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DEVELOPMENTAL BIOLOGY 1978年第1期66卷 32-42页

作者： RICHARDS, G CAMBRIDGE UNIV DEPT GENET CAMBRIDGE CB2 3EH ENGLAND

In the salivary gland chromosomes of late-3rd instar larvae and in late (8- to 12-h) prepupae of D. melanogaster, there are ecdysone-induced sequences of puffing patterns which can be reproduced in vitro. These 2 sequences are separated by a period when the glands are thought to be exposed to a low titer of .beta.-ecdysone and during which they acquire the competence to respond to ecdysone at the late prepupal puff sites. Attempts to modify either the late larval or the late prepupal responses to ecdysone in vitro by the simultaneous addition of juvenile hormone (JH) with ecdysone, to larval or prepupal glands, respectively, are unsuccessful. JH (about 10-6 M) is added to larval glands cultured 6 h in ecdysone and then 3 h in JH alone, the subsequent induction of prepupal ecdysone puffs is inhibited. The role of JH appears to lie in modifying the acquisition of competence to respond to ecdysone rather than in a direct antagonism between the 2 hormones.

关键词：染色体/超微结构培养基培养技术放线菌酮/药理学黑腹果蝇/遗传学蜕皮激素/药理学保幼激素类/药理学幼虫/遗传学涎腺/细胞学转录遗传动物

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