Analysis of the transcription unit adjacent to the 3′-end of the DOPA decarboxylase gene in Drosophila melanogaster

Ca, Spencer; Rd, Gietz; Hodgetts, RB

doi:10.1016/0012-1606(86)90402-1

Cited by 12 publications

(3 citation statements)

References 8 publications

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“…Although the larger cDNA did not contain a string of As at its 3' end, the smaller cDNA isolated from the same library did contain a string of As with the first A of the poly(A) tail at position 4590. This is in excellent agreement with the studies of Spencer et al (1986) which show that Ddc RNAs from mature larvae and young adults terminate near the HpaI site at position 4526. The polyadenylation signal AAUACA (underlined in Figure 2) is located 14 bp upstream from the polyadenylation site.…”

Section: Resultssupporting

confidence: 91%

Sequence and structure of the dopa decarboxylase gene of Drosophila: evidence for novel RNA splicing variants.

Eveleth¹,

Gietz²,

Spencer³

et al. 1986

The EMBO Journal

115

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In Drosophila, dopa decarboxylase (DDC) serves a dual role in neurotransmitter production and sclerotization of the cuticle. The Ddc gene is under complex hormonal and tissuespecific control and several sizes of Ddc RNA are observed at embryonic hatching, pupariation and adult eclosion. We present here the complete nucleotide sequence of the Drosophila dopa decarboxylase gene and the partial sequence of two corresponding Ddc cDNAs. The sequence allows us to account for the detailed structure of four of the five major Ddc RNA species observed. The cDNA sequence reveals the existence of previously undetected splicing events and provides evidence for two RNA splicing alternatives which appear to encode two protein isoforms. The structure, processing and developmental regulation of the Ddc transcripts and putative protein isoforms are discussed. Interestingly, the pyridoxal-binding peptide of porcine DDC matches the Drosophila sequence perfectly suggesting considerable selective pressure on at least portions of the sequence. This is the first available Ddc gene sequence from any organism and should serve as a basis of comparison for the related proteins of other species.

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Section: Resultssupporting

confidence: 91%

Sequence and structure of the dopa decarboxylase gene of Drosophila: evidence for novel RNA splicing variants.

Eveleth¹,

Gietz²,

Spencer³

et al. 1986

The EMBO Journal

115

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“…The Ddc transcripts detected in hormone-treated cells were approximately 500 nucleotides longer than the homologous transcripts reported by Hirsh and Davidson (19) and by Hodgetts and co-workers (16,35) The polyclonal nature of the DDC antiserum used in this study resulted in additional complexity of an already perplexing set of results. The presence of an abundant non-DDC 55-kDa protein which cross-reacts with the antiserum may explain the 55-kDa band which is detected in hormonally naive cells (Fig.…”

Section: Kineticsmentioning

confidence: 55%

“…The specific activity of 125I-labeled protein A was 30 Ci/g (ICN Pharmaceuticals Inc., Irvine, Calif.). (1,16,35) and adult flies (4.0, 3.0, and 2.0 kb; Fig. 2).…”

Section: Methodsmentioning

confidence: 99%

Modulation of novel-length DOPA decarboxylase transcripts by 20-OH-ecdysone in a Drosophila melanogaster Kc cell subline.

Swiderski¹,

O’Connor²

1986

Mol. Cell. Biol.

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The induction of DOPA decarboxylase (DDC) activity by 20-OH-ecdysone (20-OHE) in a subline of Drosophila melanogaster Kc cells was investigated. Cells cultured in the continuous presence of the steroid hormone exhibited a 96-h temporal lag prior to a peak of DDC enzyme activity while arrested in the G2 phase of the cell cycle. The concentration of Ddc RNA increased sixfold between 72 and 96 h after initial exposure to hormone. Similarly, this increase was correlated temporally with a 26-fold increase in DDC enzyme activity. The Kc Ddc primary transcript, processing intermediate, and mature mRNA all were approximtely 500 nucleotides longer than the corresponding transcripts observed for newly eclosed adult D. melanogaster. In vitro translation of poly(A)+ RNA from Kc cells resulted in an immunoprecipitable polypeptide which exhibited similar mobility on sodium dodecyl sulfate gels to that of DDC synthesized in vitro by larval epidermal poly(A)+ RNA.DOPA decarboxylase (DDC; EC 4.1.1.28) is responsible for the production of compounds which cross-link and color the insect cuticle (25). Enzyme activity is found predominantly in the epidermis (25), although minor amounts are associated with the nervous system for neurotransmitter synthesis (12,24) and with the ovary (44). The appearance of DDC during Drosophila melanogaster development is regulated in a stage-and tissue-specific manner by the insect steroid hormone 20-OH-ecdysone (20-OHE; 21, 25). Regulation of DDC activity is complex and exhibits elements of transcriptional (8,16,19,21) and posttranscriptional control (1,8).Previous studies have demonstrated the induction of DDC activity in Kc-derived cells after a 48-h lag following the administration of physiological concentrations of 20-OHE (37). In the present study, an increase in the steady-state mass of Ddc transcripts was correlated temporally with a 26-fold induction of enzyme activity. Moreover, the ecdysteroid-induced Ddc transcripts in these cells appear to be longer than the homologous sequences induced in vivo. The Kc transcripts were characterized with regard to nucleotide length and homology to several D. melanogaster Ddc gene probes. In addition, in vitro translation and immunoprecipitation were used to further characterize the Ddc gene product in Kc cells. MATERIALS AND METHODSKc cell culture and 20-OHE treatment. The D. melanogaster 7E10(4)EC subline was derived from the parental Kc line and was maintained in D-20 medium as described by Echalier and Ohanessian (13). 20-OH-Ecdysone (2pi,3 ,14a,20R,22R,25-hexahydroxy-5-cholest-7-ene-6one) was purchased from Simes, Milan, Italy. DDC activity was induced at 4 x 10-7 M 20-OHE in roller cultures having a density of 4 x 106 cells per ml. Following treatment, cells were centrifuged at 500 x g for 10 min at 4°C, resuspended in cold D-20 medium, repelleted, and frozen immediately at -700C. 15,000 x g. As a control, 10 newly eclosed flies were homogenized in 1 ml of buffer and assayed for DDC activity. A 3-,ul sample of each homogenate was incubated in tr...

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The Genetic and Molecular Organization of the Dense Cluster of Functionally Related, Vital Genes in the DOPA Decarboxylase Region of the Drosophila melanogaster Genome

Wright

1987

Results and Problems in Cell Differentiation

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Analysis of the transcription unit adjacent to the 3′-end of the DOPA decarboxylase gene in Drosophila melanogaster

Cited by 12 publications

References 8 publications

Sequence and structure of the dopa decarboxylase gene of Drosophila: evidence for novel RNA splicing variants.

Sequence and structure of the dopa decarboxylase gene of Drosophila: evidence for novel RNA splicing variants.

Modulation of novel-length DOPA decarboxylase transcripts by 20-OH-ecdysone in a Drosophila melanogaster Kc cell subline.

The Genetic and Molecular Organization of the Dense Cluster of Functionally Related, Vital Genes in the DOPA Decarboxylase Region of the Drosophila melanogaster Genome

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