Embryonic Pattern in Drosophila: The Spatial Distribution and Sequence-specific DNA Binding of engrailed Protein

O’Farrell, Patrick H.; Desplan, Claude; DiNardo, Stephen; Kassis, Judith A.; Kuner, Jerry M.; Sher, Elizabeth; Theis, James F.; Wright, Diana M.

doi:10.1101/sqb.1985.050.01.030

Cited by 8 publications

(3 citation statements)

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“…In D. melanogaster , en is a segment polarity gene expressed in segmental stripes in the primordia of the posterior compartment of each segment [ 33 , 34 ]. Similar segmental expression patterns for en were observed in a wide range of other species including all insects examined to date as well as more distant arthropods [ 35 – 38 ].…”

Section: Resultsmentioning

confidence: 99%

Establishment of molecular genetic approaches to study gene expression and function in an invasive hemipteran, Halyomorpha halys

Chen

Reding

et al. 2017

EvoDevo

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Hemiptera is a large clade of insects understudied in terms of developmental biology. Halyomorpha halys, the Brown Marmorated Stink Bug (BMSB, referred to throughout as H. halys), is an invasive hemipteran pest of the mid-Atlantic region of the USA that has rapidly spread to other regions in recent years, devastating a wide range of crops using a piercing and sucking mechanism. Its phylogenetic position, polyphagous habits, and rapid spread in the USA suggested that H. halys would be an ideal system to broaden our knowledge of developmental mechanisms in insects. We and others previously generated transcriptome sequences from different life stages of this insect. Here, we describe tools to examine gene expression patterns in whole-mount H. halys embryos and to test the response of H. halys to RNA interference (RNAi). We show that spatial and temporal patterns of gene expression in H. halys can be effectively monitored by both immunostaining and in situ hybridization. We also show that delivery of dsRNA to adult females knocks down gene function in offspring, using the homeotic gene Sex combs reduced (Scr). Knockdown of Hh-Scr resulted in dramatic malformations of the mouthparts, demonstrating for the first time that RNAi is effective in this species. Our results suggest that, despite difficulties with long-term laboratory culture of H. halys, this species shows promise as a developmental system.Electronic supplementary materialThe online version of this article (doi:10.1186/s13227-017-0078-6) contains supplementary material, which is available to authorized users.

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Section: Resultsmentioning

confidence: 99%

Establishment of molecular genetic approaches to study gene expression and function in an invasive hemipteran, Halyomorpha halys

Chen

Reding

et al. 2017

EvoDevo

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“…For example, en is first expressed in a series of stripes which transect the anteriorposterior axis of the embryo (11,19,33). Later in development, en is expressed in particular cells in the head, terminalia, central and peripheral nervous systems (39), and imaginal disks (8,33). Molecular mapping of mutations of the en gene and other developmental loci having intricate spatial patterns of expression has shown that very large cis-acting regulatory regions are associated with these genes.…”

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confidence: 99%

Evolutionary conservation of homeodomain-binding sites and other sequences upstream and within the major transcription unit of the Drosophila segmentation gene engrailed.

Kassis¹,

Desplan²,

Wright³

et al. 1989

Mol. Cell. Biol.

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The engrailed (en) gene functions throughout Drosophila development and is expressed in a succession of intricate spatial patterns as development proceeds. Normal en function relies on an extremely large cis-acting regulatory region (70 kilobases). We are using evolutionary conservation to help identify en sequences important in regulating patterned expression. Sequence comparison of 2.6 kilobases upstream of the en coding region of D. melanogaster and D. virilis (estimated divergence time, 60 million years) showed that 30% of this DNA occurs in islands of near perfect sequence conservation. One of these conserved islands contains binding sites for homeodomain-containing proteins. It has been shown genetically that homeodomain-containing proteins regulate en expression. Our data suggested that this regulation may be direct. The remaining conserved islands may contain binding sites for other regulatory proteins.Many of the genes that control development in Drosophila have been identified by mutation and analyzed genetically and molecularly. Studies of the action and interaction of these genes have led to their classification into a hierarchy (for reviews, see references 26 and 49). The patterns of expression of developmentally important genes in Drosophila can be extremely complex. For example, en is first expressed in a series of stripes which transect the anteriorposterior axis of the embryo (11,19,33). Later in development, en is expressed in particular cells in the head, terminalia, central and peripheral nervous systems (39), and imaginal disks (8, 33). Molecular mapping of mutations of the en gene and other developmental loci having intricate spatial patterns of expression has shown that very large cis-acting regulatory regions are associated with these genes. For example, the en gene extends across 70 kilobases (kb) but has only a 4.5-kb transcription unit (15,34). Using in vitro DNA constructs and P-element transformation, we have begun to functionally dissect the en cis-acting regulatory regions (13; J. A. Kassis, manuscript in preparation). Because of the large size of the cis-regulatory region, we are guiding our search for important regulatory sequences by identifying sequences that are evolutionarily conserved.We isolated the en gene from a distantly related Drosophila species, D. virilis, and compared it with the D. melanogaster gene (28,29). Importantly, these two species are sufficiently separated such that only functional sequences should be conserved (estimated divergence time, 60 million years [2]). Also important is that en expression is similar in the two species (J. A. Kassis, unpublished data). Here 3.1 kb from D. virilis. We found that about 30% of this DNA is conserved between the two species. The conservation is best described as islands of near perfect conservation within largely divergent regions. We showed that some of the conserved DNA sequences contain in vitro binding sites for homeodomain (HD)-containing proteins. Mutations in genes which encode HD-containing proteins alter en e...

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“…Since the mutant possessed a dent in the scutellum Eker called it Engrailed after ‘engrailé’ a heraldic term from middle-age French meaning ‘dented by hail’. Engrailed turned out to be a key selector gene that is involved in the development of posterior compartments of appendages and segments [ 14 – 16 ] and the nervous system [ 17 , 18 ] during Drosophila development. Since then, one or more Engrailed proteins have been described in many metazoans from echinoderms [ 19 ], nematodes [ 20 ], annelids [ 21 ], brachiopods [ 22 ], platyhelminthes [ 23 ], molluscs [ 24 ], cephalochordates [ 25 ], onychophorans [ 26 ] priapulids [ 27 ] and in vertebrates [ 28 ].…”

Section: Introductionmentioning

confidence: 99%

Engrailed homeoproteins in visual system development

Wizenmann

Stettler

Moya

2014

Cell. Mol. Life Sci.

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Engrailed is a homeoprotein transcription factor. This family of transcription factors is characterized by their DNA-binding homeodomain and some members, including Engrailed, can transfer between cells and regulate protein translation in addition to gene transcription. Engrailed is intimately involved in the development of the vertebrate visual system. Early expression of Engrailed in dorsal mesencephalon contributes to the development and organization of a visual structure, the optic tectum/superior colliculus. This structure is an important target for retinal ganglion cell axons that carry visual information from the retina. Engrailed regulates the expression of Ephrin axon guidance cues in the tectum/superior colliculus. More recently it has been reported that Engrailed itself acts as an axon guidance cue in synergy with the Ephrin system and is proposed to enhance retinal topographic precision.

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Embryonic Pattern in Drosophila: The Spatial Distribution and Sequence-specific DNA Binding of engrailed Protein

Cited by 8 publications

References 0 publications

Establishment of molecular genetic approaches to study gene expression and function in an invasive hemipteran, Halyomorpha halys

Establishment of molecular genetic approaches to study gene expression and function in an invasive hemipteran, Halyomorpha halys

Evolutionary conservation of homeodomain-binding sites and other sequences upstream and within the major transcription unit of the Drosophila segmentation gene engrailed.

Engrailed homeoproteins in visual system development

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