Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions

Ingolia, Thomas D.

doi:10.1016/0092-8674(80)90430-4

Cited by 287 publications

(163 citation statements)

References 31 publications

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“…Sites for restriction enzymes are represented as follows: (B) XbaI; (H3) HindlII; {N) NotI; (X)XhoI. codon in this minigene, beginning at nucleotide -205 in the hsp70 promoter (Ingolia et al 1980), was GCGAAAGCTAAG-CAAATAGGATCCGTTAACGAA(ATG). Transgenic fly lines containing the hsro and sevro transcriptional fusions were produced by P-element-mediated transformation (Spradling and Rubin 1982).…”

Section: Transgenic Fly Linesmentioning

confidence: 99%

“…Thus, a HindIII/BglII digestion of this plasmid released the 5' 717 bp of the repaired eDNA. This fragment was purified and ligated in a three-way ligation to a 470-bp EcoRI-HindIII fragment of the heat shock promoter (which corresponds to the XbaI-XmnI fragment of Ingolia et al 1980) and a BglII-EcoRI fragment containing the 3' 1402 bp of the rough genomic rescue fragment ). The resulting Bluescript plasmid, pbshsro, has a heat shock promoter transcriptionally fused to a rough minigene missing the first intron but containing the second intron and 3'-untranslated region, including the poly(A) site of the genomic rough gene (see Fig.…”

Section: Transgenic Fly Linesmentioning

confidence: 99%

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The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype.

Kimmel¹,

Heberlein²,

Rubin³

1990

Genes Dev.

263

131

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The Drosophila homeo box gene rough is required in photoreceptor cells R2 and R5 for normal eye development. We show here that rough protein expression is limited to a subset of cells in the developing retina where it is transiently expressed for 30--60 hr. The rough protein is first expressed broadly in the morphogenetic furrow but is rapidly restricted to the R2, R3, R4, and RS precursor cells. Ubiquitous expression of rough under the control of the hspTO promoter in third-instar larvae supresses the initial steps of ommatidial assembly.Structures derived from other imaginal discs are not affected. Ectopic expression of rough in the R7 precursor, through the use of the sevenless promoter, causes this cell to develop into an R1-6 photoreceptor subtype; however, this cell still requires sevenless function for its neural differentiation. Taken together with previous analyses of the rough mutant phenotype, these results suggest that the normal role of rough is to establish the unique cell identity of photoreceptors R2 and R5.[Key Words: Homeo domain; eye development; Drosophila melanogaster; rough; ectopic expression; sevenless] Received January 9, 1990; revised version accepted February 21, 1990.The Drosophila eye is composed of -800 unit eyes called ommatidia. Each ommatidium contains a stereotyped arrangement of 20 cells in which each cell can be identified by its morphology and position (Dietrich 1909;Ready et al. 1976). Eight of these cells are photoreceptors: six outer cells, R1-R6, and two smaller central cells, R7 and R8. The adult retina develops from an undifferentiated epithelial monolayer, the eye imaginal disc, beginning in the third instar larva (Ready et al. 1976; Tomlinson and Ready 1987aj. Organization of cells into ommatidia begins in an indentation in the disc, called the morphogenetic furrow, which moves from posterior to anterior across the eye disc over a 2-day period. Thus, ommatidial assembly occurs asynchronously across the anterior-posterior axis of the eye disc such that each column of developing ommatidia is -2 hr more developmentally advanced than the adjacent column to the anterior. In the morphogenetic furrow, five cells form a circular precluster that eventually gives rise to five of the eight photoreceptor cells of an ommatidium. Neuronal differentiation of these five photoreceptor precursors occurs in a stereotyped order: R8, R2, and R5, and R3 and R4 (Tomlinson and Ready 1987a An eye disc from a larva homozygous for the null rough allele ro x63 stained with MAbro 1. ro ~63 is a small X-ray-induced deletion near the 5' end of the rough-coding region that results in a frameshift after the thirty-first codon {U. Heberlein, unpubl.). The lack of detectable rough protein in this disc demonstrates the specificity of MAbrol. The arrowhead in A marks the position of the morphogenetic furrow where rough protein was first detected, rough protein was localized in the cell nucleus. Magnifications in A, A inset, and B are 250 x, 350 x, and 200 x, respectively. Phase-contrast images of singl...

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Section: Transgenic Fly Linesmentioning

confidence: 99%

Section: Transgenic Fly Linesmentioning

confidence: 99%

The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype.

Kimmel¹,

Heberlein²,

Rubin³

1990

Genes Dev.

263

131

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“…These four genes are clustered within an 1,1 kb region, but are not all transcribed in the same direction; each is transcribed into a separate mRNA (11,12). This paper reports an extension of this characterization and a determination of the primary sequence of the 5' flanking regions of these four genes, as well as a comparison with the 5' flanking regions of the major hsp70 heat shock genes (13) MATERIALS AND …”

Section: Introductionmentioning

confidence: 99%

Primary sequence of the 5′ flanking regions of the Drosophila heat shock genes in chromosome subdivision 67B

1981

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The 5' flanking regions of the four small heat shock genes of Drosophila melanogaster from cytological locus 67B have been characterized. Approximately 500 bp of the primary sequence upstream from the proposed site of initiation of translation has been determined and the 5' end of the messenger RNAs have been localized for each gene.Each of the four genes contains an A-T rich sequence, either TATAAATA or TATAAAAG, which is flanked by a G-C rich region. This A-T rich sequence, which ends about 23 bp upstream from the proposed site of initiation of transcription, is similar to those found in most eukaryotic genes. Novel features of these four genes include a region of homology beginning near the proposed site of initiation of transcription and extending about 20 bp into the 5' noncoding region of the genes. This sequence is also found in the gene for the major heat shock protein, hsp 70. The leaders of these five heat shock genes are long, from 111 to 253 bases in length, as well as unusually A rich, from 46% to 51% A. In addition, each of the four small genes contains the sequence ACTTTNA, 195+12 bp from the proposed site of initiation of transcrip tion.

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“…As all components used in these constructions have been sequenced, the entire nucleotide sequence of the Icarus element can be predicted and is available on request. Below we list the origin of the various constituents of Icarus, starting with the 5' inverted repeat of the P element: Icarus nucleotides (nt) 1 through 732 correspond to nt 1 through 732 from p6.1 (29); nt 733 through 3383 are from pUC9 (44); nt 3384 through 3837 contain the hsp7O promoter (17,37); nt 3838 through 3858 are pUC9 polylinker sequences; nt 3859 through 6700 correspond to nucleotides 39 through 2880 from pur25.1 (29); nt 6701 through 6710 come from the pUC9 polylinker (44); nt 6711 through 7099 correspond to nt 729 through 893 and 2686 through 2907 of the P element (29); and nt 7100 through 8622 come from the white gene (28).…”

Section: Methodsmentioning

confidence: 99%

P transposons controlled by the heat shock promoter.

Steller¹,

Pirrotta²

1986

Mol. Cell. Biol.

104

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We have transformed Drosophila melanogaster with modified P-element transposons, which express the transposase function from the heat-inducible hsp70 heat shock promoter. The Icarus transposon, which contains a direct hsp7O-P fusion gene, behaved like a very active autonomous P element even before heat shock induction. Although heat shock led to abundant somatic transcription, transposition of the Icarus element was confined to germ line cells. To reduce the constitutive transposase activity observed for the Icarus element, we attenuated the translational efficiency of the transposase RNA by inserting the transposon 5 neomycin resistance gene between the hsp70 promoter and the P-element sequences. The resulting construct, called Icarus-neo, conferred resistance to G418, and its transposition was significantly stimulated by heat shock. Heat shocks applied during the embryonic or third instar larval stage had similar effects, indicating that transposition of P elements is not restricted to a certain developmental stage. Both Icarus and Icarus-neo destabilized snW in a P-cytotype background and thus at least partially overcome the repression of transposition. Our results suggest that the regulation of P-element transposition occurs at both the transcriptional and posttranscriptional levels.Transposable elements are a major cause of genome instability, responsible for a large fraction of spontaneous mutations in Drosophila melanogaster (1,3,7,20,25,26,31,43). The functional analysis of metazoan transposons has been complicated by the fact that transposition events are usually rare and the physiological conditions they require are not well defined. The P elements responsible for the phenomenon of P-M hybrid dysgenesis (21) in D. melanogaster are of particular interest because their transpositional activity, which can be readily detected by genetic tests (14,19,36), is potentially high but under strict genetic control. Transposition is induced at a high frequency only if males carrying functional P elements (P strains) are crossed with females lacking P factors (M strains), but not in the reciprocal or in a P x P cross (for a review, see references 6 and 13). Because the repressed state (P cytotype) is determined by P factors themselves, it has been suggested that P elements code for a negative regulator in addition to transposase function (10, 29).Transposition of P elements is specific for germ line cells since no appreciable activity can be detected in the soma (11,42). It is not known whether this is the result of tissuespecific transcription of P elements or a requirement for germ line-specific host functions acting posttranscriptionally.P elements have been cloned (4, 32) and sequenced (29) and shown to transpose from injected plasmid DNA into the chromosomes of germ line cells (19,33,36 elements can no longer transpose autonomously, they can be mobilized when the missing transposase function is provided in trans by an intact helper element. A functional analysis of P elements was started by Karess and Rubin (1...

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Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions

Cited by 287 publications

References 31 publications

The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype.

The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype.

Primary sequence of the 5′ flanking regions of the Drosophila heat shock genes in chromosome subdivision 67B

P transposons controlled by the heat shock promoter.

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