scholarly journals Analysis of the Promoter of the ninaE Opsin Gene in Drosophila melanogaster

Genetics ◽  
1987 ◽  
Vol 116 (4) ◽  
pp. 565-578 ◽  
Author(s):  
Drzislav Mismer ◽  
Gerald M Rubin
Keyword(s):  
Control Region ◽  
Photoreceptor Cells ◽  
P Element ◽  
Opsin Gene ◽  
Regulatory Sequences ◽  
Enhancer Element ◽  
Cis Acting ◽  
E Coli ◽  
Promoter Function ◽  
Control Regions

ABSTRACT We have analyzed the cis-acting regulatory sequences of the ninaE gene. This gene encodes the major Drosophila melanogasteropsin, the protein component of the primary chromophore of photoreceptor cells R1-R6 of the adult eye. DNA fragments containing the start point of transcription of the ninaE gene were fused to either the Escherichia coli chloramphenicol acetyltransferase or lacZ (β-galactosidase) gene and introduced into the Drosophila germline by P-element-mediated transformation. Expression of the E. coli genes was then used to assay the ability of various sequences from the ninaE gene to confer the ninaE pattern of expression. Fragments containing between 2.8 kb and 215 bp of the sequences upstream of the start of transcription plus the first 67 bp of the untranslated leader were able to direct nearly wild-type expression. We have identified three separable control regions in the ninaE promoter. The first, which has the properties of an enhancer element, is located between nucleotides -501 and -219. The removal of this sequence had little effect on promoter function; this sequence appears to be redundant. However, it appears to be able to substitute for the second control region which is located between nucleotides -215 and -162, and which also affects the level of output from this promoter. Removal of these two control regions resulted in a 30-fold decrease in expression; however tissue specificity was not affected. The third control region, located downstream from nucleotide -120, appears to be absolutely necessary for promoter function in the absence of the first two regulatory sequences. Examination of larvae containing fusion genes expressing β-galactosidase suggests that the ninaE gene is also expressed in a subset of cells in the larval photoreceptor organ.

scholarly journals Definition of cis-acting elements regulating expression of the Drosophila melanogaster ninaE opsin gene by oligonucleotide-directed mutagenesis.

Genetics ◽  
1989 ◽  
Vol 121 (1) ◽  
pp. 77-87 ◽  
Author(s):  
D Mismer ◽  
G M Rubin
Keyword(s):  
Drosophila Melanogaster ◽  
The Body ◽  
Drosophila Virilis ◽  
P Element ◽  
Opsin Gene ◽  
Directed Mutagenesis ◽  
Homologous Gene ◽  
Regulatory Sequences ◽  
Specific Expression ◽  
Cis Acting

Abstract We have analyzed the cis-acting regulatory sequences of the Rh1 (ninaE) gene in Drosophila melanogaster by P-element-mediated germline transformation of indicator genes transcribed from mutant ninaE promoter sequences. We have previously shown that a 200-bp region extending from -120 to +67 relative to the transcription start site is sufficient to obtain eye-specific expression from the ninaE promoter. In the present study, 22 different 4-13-bp sequences in the -120/+67 promoter region were altered by oligonucleotide-directed mutagenesis. Several of these sequences were found to be required for proper promoter function; two of these are conserved in the promoter of the homologous gene isolated from the related species Drosophila virilis. Alteration of a conserved 9-bp sequence results in aberrant, low level expression in the body. Alteration of a separate 11-bp sequence, found in the promoter regions of several photoreceptor-specific genes of Drosophila, results in an approximately 15-fold reduction in promoter efficiency but without apparent alteration of tissue-specificity. A protein factor capable of interacting with this 11-bp sequence has been detected by DNaseI footprinting in embryonic nuclear extracts. Finally, we have further characterized two separable enhancer sequences previously shown to be required for normal levels of expression from this promoter.

Analysis of the promoter of the Rh2 opsin gene in Drosophila melanogaster.

Genetics ◽  
1988 ◽  
Vol 120 (1) ◽  
pp. 173-180
Author(s):  
D Mismer ◽  
W M Michael ◽  
T R Laverty ◽  
G M Rubin
Keyword(s):  
Drosophila Melanogaster ◽  
Fusion Gene ◽  
Photoreceptor Cells ◽  
Histochemical Staining ◽  
P Element ◽  
Opsin Gene ◽  
Regulatory Sequences ◽  
Specific Expression ◽  
Beta Galactosidase ◽  
Cat Expression

Abstract We have analyzed the cis-acting regulatory sequences of the Drosophila melanogaster Rh2 gene that encodes the protein component of a rhodopsin which is expressed in ocellar photoreceptor cells. DNA fragments containing the start point of transcription of the Rh2 gene were fused to either the Escherichia coli chloramphenicol acetyltransferase (CAT) or lacZ (beta-galactosidase) genes and introduced into the Drosophila germline by P-element-mediated transformation. Expression of the E. coli genes was then used to assay the ability of various sequences from the Rh2 gene to confer upon the indicator genes the Rh2 pattern of expression. Fragments containing between 4.3 kb and 183 bp upstream of the start of transcription plus the first 32 bp of the 5'-untranslated leader were found to result in nearly identical levels of head-specific CAT expression. Deletion of Rh2 sequences distal to position -112 bp resulted in loss of detectable CAT expression from these Rh2/CAT fusion constructs. We have, therefore, defined a region essential for head-specific expression of the Rh2 gene to a region extending from -183 to -112. We have determined the DNA sequence of the Rh2 promoter from -448 to +32 and have found an 11-bp sequence which is also present in the upstream flanking sequences of two other photoreceptor-specific genes (ninaE and ninaC). By histochemical staining of beta-galactosidase expressed under the control of the Rh2 promoter and by analyzing the effect of the ocelliless mutation on the expression of an Rh2/CAT fusion gene, we have been able to demonstrate that this promoter is active in ocelli.

Target sequences for hunchback in a control region conferring Ultrabithorax expression boundaries

Development ◽  
1991 ◽  
Vol 113 (4) ◽  
pp. 1171-1179 ◽  
Author(s):  
C.C. Zhang ◽  
J. Muller ◽  
M. Hoch ◽  
H. Jackle ◽  
M. Bienz
Keyword(s):  
Long Range ◽  
Control Region ◽  
Binding Sites ◽  
Regulatory Sequences ◽  
Stripe Pattern ◽  
Anteroposterior Axis ◽  
Target Sequences ◽  
Beta Galactosidase ◽  
Control Regions

Boundaries of Ultrabithorax expression are mediated by long-range repression acting through the PBX or ABX control region. We show here that either of these control regions confers an early band of beta-galactosidase expression which is restricted along the anteroposterior axis of the blastoderm embryo. This band is succeeded by a stripe pattern with very similar anteroposterior limits. Dissection of the PBX control region demonstrates that the two patterns are conferred by distinct cis-regulatory sequences contained within separate PBX subfragments. We find several binding sites for hunchback protein within both PBX subfragments. Zygotic hunchback function is required to prevent ectopic PBX expression. Moreover, the PBX pattern is completely suppressed in embryos containing uniformly distributed maternal hunchback protein. Our results strongly suggest that hunchback protein directly binds to the PBX control region and acts as a repressor to specify the boundary positions of the PBX pattern.

Multiple hormone-inducible enhancers as mediators of differential transcription

1986 ◽  
Vol 6 (12) ◽  
pp. 4526-4538
Author(s):  
M G Toohey ◽  
K L Morley ◽  
D O Peterson
Keyword(s):  
Regulatory Control ◽  
Regulatory Sequences ◽  
Enhancer Element ◽  
Cis Acting ◽  
Mouse Mammary Tumor ◽  
Tumor Virus ◽  
Differential Transcription ◽  
Virus Promoter ◽  
Inducible Gene

Sets of genes under a common regulatory control in a given cell type are often differentially transcribed. The possibility that this differential transcription can be modulated by the number or strength of cis-acting regulatory sequences associated with a given gene was tested by using the glucocorticoid-responsive enhancer element associated with the mouse mammary tumor virus promoter. Results indicate that differential levels of hormone-inducible gene expression can be modulated in an additive way by the number of glucocorticoid-responsive enhancers associated with this promoter. Realization of these effects shows little preference for position of the additional elements with respect to the promoter. When sequences that bind the glucocorticoid receptor in vitro with somewhat lower affinity than the enhancer were tested, these additive effects were not detected. The results support that differential transcription of genes subject to a common regulatory control can be mediated, at least in part, by the number or strength of their associated cis-acting regulatory sequences.

Cis-acting regulatory sequences governing Wnt-1 expression in the developing mouse CNS

Development ◽  
1994 ◽  
Vol 120 (8) ◽  
pp. 2213-2224 ◽  
Author(s):  
Y. Echelard ◽  
G. Vassileva ◽  
A.P. McMahon
Keyword(s):  
Neural Crest ◽  
De Novo ◽  
Ectopic Expression ◽  
Neural Crest Cells ◽  
Neural Tube Closure ◽  
Regulatory Sequences ◽  
Lacz Gene ◽  
Mouse Development ◽  
Enhancer Element ◽  
Cis Acting

The protooncogene Wnt-1 encodes a short-range signal which is first expressed in, and appears to demarcate, the presumptive midbrain. Absence of Wnt-1 expression leads to the loss of this region of the brain. By the end of neural tube closure, expression of Wnt-1 extends down much of the dorsal midline of the central nervous system (CNS). Expression is exclusively limited to the CNS at this and later stages. We have investigated the regulation of Wnt-1 during mouse development. Analysis of the embryonic expression of Wnt-1-lacZ reporter constructs spanning nearly 30 kb of the Wnt-1 locus identified a 5.5 kb cis-acting 3′ enhancer element which confers correct temporal and spatial expression on the lacZ gene. Interestingly embryos express Wnt-1-lacZ transgenes in migrating neural crest cells which are derived from the dorsal CNS. Ectopic expression of the Wnt-1-lacZ transgenes may result from perdurance of beta-galactosidase activity in migrating neural crest cells originating from a Wnt-1-expressing region of the dorsal CNS. Alternatively, ectopic expression may arise from transient de novo activation of the transgenes in this cell population. These results are a first step towards addressing how regional cell signaling is established in the mammalian CNS. In addition, transgene expression provides a new tool for the analysis of neural crest development in normal and mutant mouse embryos.

scholarly journals Multiple hormone-inducible enhancers as mediators of differential transcription.

1986 ◽  
Vol 6 (12) ◽  
pp. 4526-4538 ◽  
Author(s):  
M G Toohey ◽  
K L Morley ◽  
D O Peterson
Keyword(s):  
Regulatory Control ◽  
Regulatory Sequences ◽  
Enhancer Element ◽  
Cis Acting ◽  
Mouse Mammary Tumor ◽  
Tumor Virus ◽  
Differential Transcription ◽  
Virus Promoter ◽  
Inducible Gene

Sets of genes under a common regulatory control in a given cell type are often differentially transcribed. The possibility that this differential transcription can be modulated by the number or strength of cis-acting regulatory sequences associated with a given gene was tested by using the glucocorticoid-responsive enhancer element associated with the mouse mammary tumor virus promoter. Results indicate that differential levels of hormone-inducible gene expression can be modulated in an additive way by the number of glucocorticoid-responsive enhancers associated with this promoter. Realization of these effects shows little preference for position of the additional elements with respect to the promoter. When sequences that bind the glucocorticoid receptor in vitro with somewhat lower affinity than the enhancer were tested, these additive effects were not detected. The results support that differential transcription of genes subject to a common regulatory control can be mediated, at least in part, by the number or strength of their associated cis-acting regulatory sequences.

scholarly journals Ingrowth by photoreceptor axons induces transcription of a retrotransposon in the developing Drosophila brain

Development ◽  
1994 ◽  
Vol 120 (5) ◽  
pp. 1049-1058 ◽  
Author(s):  
B.A. Mozer ◽  
S. Benzer
Keyword(s):  
P Element ◽  
Regulatory Sequences ◽  
Lacz Expression ◽  
Specific Expression ◽  
Enhancer Element ◽  
Drosophila Brain ◽  
First Optic Ganglion ◽  
Combined Action ◽  
Eye Imaginal Disc ◽  
The Brain

The development of the lamina, the first optic ganglion of the fly visual system, depends on inductive cues from the innervating photoreceptor axons. lacZ expression from a P-element insertion, A72, occurs in the anlage of the lamina coincident with axon ingrowth from the eye imaginal disc. In eyeless mutants lacking photoreceptor axons, lacZ expression did not occur. The P-element was found to have inserted within the 3′ long terminal repeat (LTR) of a ‘17.6′ type retrotransposon. The expression pattern of 17.6 transcripts in the brain in wild-type and eyeless mutants paralleled the expression of the lacZ reporter. Analysis of 17.6 cis-regulatory sequences indicates that the lamina-specific expression is due to the combined action of an enhancer element in the LTR and a repressor element within the internal body of the retrotransposon. The regulation of the 17.6 retrotransposon provides a model for the study of innervation-dependent gene expression in postsynaptic cells during neurogenesis.

scholarly journals Activation of gene expression by adenovirus and herpesvirus regulatory genes acting in trans and by a cis-acting adenovirus enhancer element

Cell ◽  
1983 ◽  
Vol 35 (1) ◽  
pp. 127-136 ◽  
Author(s):  
Michael J. Imperiale ◽  
Lawrence T. Feldman ◽  
Joseph R. Nevins
Keyword(s):  
Gene Expression ◽  
Regulatory Genes ◽  
Enhancer Element ◽  
Cis Acting ◽  
In Trans

Reconstitution of the vitamin D-responsive osteocalcin transcription unit in Saccharomyces cerevisiae

1989 ◽  
Vol 9 (8) ◽  
pp. 3517-3523
Author(s):  
D P McDonnell ◽  
J W Pike ◽  
D J Drutz ◽  
T R Butt ◽  
B W O'Malley
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Vitamin D ◽  
Transcription Factors ◽  
Promoter Activity ◽  
Transcription Unit ◽  
Proximal Promoter ◽  
Regulatory Sequences ◽  
Cis Acting ◽  
Osteocalcin Gene ◽  
Definition Of

The human osteocalcin gene is regulated in mammalian osteoblasts by 1,25(OH)2D3-dependent and -independent mechanisms. The sequences responsible for this activity have been mapped to within the -1339 region of the gene. We show here that this enhancer region functions analogously in Saccharomyces cerevisiae cells engineered to produce active 1,25(OH)2D3 receptor. When fused to the proximal promoter elements of the yeast iso-1-cytochrome c gene, the enhancer demonstrated substantial promoter activity. This activity was elevated further by 1,25(OH)2D3 when the reporter constructs were assayed in cells containing the 1,25(OH)2D3 receptor. This system affords a model for 1,25(OH)2D3 action and represents a simple assay system that will enable definition of the important cis-acting regulatory sequences within the osteocalcin gene and identification of their cognate transcription factors.

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