Molecular and genetic characterization of the Drosophila melanogaster 87E actin gene region.

Abstract A combined molecular and genetic analysis of the 87E actin gene (Act87E) in Drosophila melanogaster was undertaken. A clone of Act87E was isolated and characterized. The Act87E transcription unit is 1.57 kb and includes a 556-base intervening sequence in the 5' leader of the gene. The protein-coding region is contiguous and encodes a protein that is greater than 93% identical to the other Drosophila actins. By in situ hybridization with a series of deficiencies that break in 87E, Act87E was localized to a region encompassing one to three faint, polytene chromosome bands. The region between the deficiency endpoints that flank the actin gene was isolated and measures approximately 24-30 kb. The closest proximal deficiency endpoint lies 8-10 kb 5' to the actin gene; the closest distal deficiency endpoint lies 16-20 kb 3' to the actin gene. A single, recessive lethal complementation group lies between the deficiency endpoints that flank the actin gene. An EMS mutagenesis screen produced four additional members of this recessive lethal complementation group. Molecular analysis of the members of this complementation group indicated that two of the newly induced mutations have deletions of approximately 1 kb in a transcribed region 4-5 kb 3' (distal) to the actin gene. This result suggests that the recessive lethal complementation group represents a gene separate from and distal to the actin gene. The mutagenesis screen failed to identify additional recessive lethal complementation groups in the actin gene-containing region. The implications of the failure to identify recessive lethal mutations in the actin gene are discussed in reference to studies of other conserved multigene families and other muscle protein mutations.

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Structure of transcripts from the homeotic Antennapedia gene of Drosophila melanogaster: two promoters control the major protein-coding region.

Molecular and Cellular Biology ◽

10.1128/mcb.6.12.4676 ◽

1986 ◽

Vol 6 (12) ◽

pp. 4676-4689 ◽

Cited By ~ 46

Author(s):

A Laughon ◽

A M Boulet ◽

J R Bermingham ◽

R A Laymon ◽

M P Scott

Keyword(s):

Drosophila Melanogaster ◽

Alternative Polyadenylation ◽

Transcription Unit ◽

Cdna Clones ◽

Major Protein ◽

Coding Region ◽

Developmentally Regulated ◽

Protein Coding ◽

Reading Frame ◽

C Terminus

The Antennapedia (Antp) homeotic gene of Drosophila melanogaster regulates segmental identity in the thorax. Loss of Antp function results in altered development of the embryonic thoracic segments or can cause legs to be transformed into antennae. Certain combinations of Antp recessive lethal alleles complement to permit normal development. The structure of the Antp gene, analyzed by sequencing cDNA clones and exons and by transcript mapping, revealed some of the basis for its genetic complexity. It has two promoters governing two nested transcription units, one unit 36 and one 103 kilobase pairs (kb) long. Both units incorporated the same protein-coding exons, all of which are located in the 3'-most 13 kb of the gene. The two promoters resulted in the attachment of either of two long noncoding leader sequences (1.5 and 1.7 kb) to a 1.1-kb open reading frame. Both transcription units used the same pair of alternative polyadenylation sites 1.4 kb apart; the choice of sites was developmentally regulated. Some of the mutations that disrupt the larger transcription unit complemented a mutation affecting the smaller one. Dominant mutations that transform antennae into legs split the gene but left the coding exons intact. The encoded protein has unusually long runs of glutamine and a homeodomain near the C terminus.

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Structure of transcripts from the homeotic Antennapedia gene of Drosophila melanogaster: two promoters control the major protein-coding region

Molecular and Cellular Biology ◽

10.1128/mcb.6.12.4676-4689.1986 ◽

1986 ◽

Vol 6 (12) ◽

pp. 4676-4689

Author(s):

A Laughon ◽

A M Boulet ◽

J R Bermingham ◽

R A Laymon ◽

M P Scott

Keyword(s):

Drosophila Melanogaster ◽

Alternative Polyadenylation ◽

Transcription Unit ◽

Cdna Clones ◽

Major Protein ◽

Coding Region ◽

Developmentally Regulated ◽

Protein Coding ◽

Reading Frame ◽

C Terminus

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Cytogenetic and complementation analyses of recessive lethal mutations induced in the X chromosome of Drosophila by three alkylating agents

Genetics Research ◽

10.1017/s0016672300015020 ◽

1974 ◽

Vol 24 (1) ◽

pp. 1-10 ◽

Cited By ~ 32

Author(s):

J. K. Lim ◽

L. A. Snyder

Keyword(s):

Drosophila Melanogaster ◽

Salivary Gland ◽

X Chromosome ◽

Alkylating Agents ◽

Complementation Analysis ◽

Ethyl Methanesulphonate ◽

Induced Mutations ◽

Recessive Lethal ◽

Lethal Mutations

SUMMARYSalivary-gland chromosomes of 54 methyl methanesulphonate- and 50 triethylene melamine-induced X-chromosome recessive lethals in Drosophila melanogaster were analysed. Two of the lethals induced by the mono-functional agent and 11 of those induced by the polyfunctional agent were found to be associated with detectable aberrations. A complementation analysis was also done on 82 ethyl methanesulphonate- and 34 triethylene melamine-induced recessive lethals in the zeste-white region of the X chromosome. The EMS-induced lethals were found to represent lesions affecting only single cistrons. Each of the 14 cistrons in the region known to mutate to a lethal state was represented by mutant alleles, but in widely different frequencies. Seven of the TEM-induced lethals were associated with deletions, only one of which had both breakpoints within the mapped region. Twenty-six of the 27 mutations in which only single cistrons were affected were mapped to 7 of the 14 known loci. One TEM- and two EMS-induced mutations were alleles representing a previously undetected locus in the zeste-white region.

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Structure and tissue-specific expression of the Drosophila melanogaster organellar-type Ca2+-ATPase gene

Biochemical Journal ◽

10.1042/bj3100757 ◽

1995 ◽

Vol 310 (3) ◽

pp. 757-763 ◽

Cited By ~ 18

Author(s):

A Magyar ◽

E Bakos ◽

A Váradi

Keyword(s):

Drosophila Melanogaster ◽

Transcription Initiation ◽

Initiation Site ◽

Drosophila Genome ◽

Coding Region ◽

Specific Expression ◽

S1 Nuclease ◽

Protein Coding ◽

Atpase Gene ◽

High Level

A 14 kb genomic clone covering the organellar-type Ca(2+)-ATPase gene of Drosophila melanogaster has been isolated and characterized. The sequence of a 7132 bp region extending from 1.1 kb 5′ upstream of the initiation ATG codon over the polyadenylation signal at the 3′ end has been determined. The gene consists of nine exons including one with an exceptional size of 2172 bp representing 72% of the protein coding region. Introns are relatively small (< 100 bp) except for the 3′ intron which has a size of 2239 bp, an exceptionally large size among Drosophila introns. Five of the introns are in the same positions in Drosophila, Artemia and rabbit SERCA1 Ca(2+)-ATPase genes. There is only one organellar-type Ca(2+)-ATPase gene in the Drosophila genome, as was shown by Southern-blot analysis [Váradi, Gilmore-Hebert and Benz (1989) FEBS Lett. 258, 203-207] and by chromosomal localization [Magyar and Váradi (1990) Biochem. Biophys. Res. Commun. 173, 872-877]. Primer extension and S1-nuclease assays revealed a potential transcription initiation site 876 bp upstream of the translation initiation ATG with a TATA-box 23 bp upstream of this site. Analysis of the 5′ region of the Drosophila organellar-type Ca(2+)-ATPase gene suggests the presence of potential recognition sequences of various muscle-specific transcription factors and shows a region with remarkable similarity to that in the rabbit SERCA2 gene. The tissue distribution of expression of the organellar-type Ca(2+)-ATPase gene has been studied by in situ RNA-RNA hybridization on microscopic sections. A low mRNA abundance can be detected in each tissue of adult flies, suggesting a housekeeping function for the gene. On the other hand a pronounced tissue specificity of expression has also been found as the organellar-type Ca(2+)-ATPase is expressed at a very high level in cell bodies of the central nervous system and in various muscles.

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Genetic structure of the abd-A gene of Drosophila

Development ◽

10.1242/dev.107.3.575 ◽

1989 ◽

Vol 107 (3) ◽

pp. 575-583 ◽

Cited By ~ 1

Author(s):

A. Busturia ◽

J. Casanova ◽

E. Sanchez-Herrero ◽

R. Gonzalez ◽

G. Morata

Keyword(s):

Genetic Structure ◽

Regulatory Region ◽

Regulatory Elements ◽

Transcription Unit ◽

Bithorax Complex ◽

Coding Region ◽

Regulatory Regions ◽

Protein Coding ◽

Spatial Regulation ◽

Correct Level

We report the embryonic and adult phenotypes of a number of mutations of the abd-A gene of the bithorax complex. Some of them result in loss of abd-A function in the whole abd-A domain and are usually lethal. These probably eliminate or inactivate abd-A protein products. Other mutations affect only part of the abd-A domain. These are viable, appear to map outside the abd-A transcription unit, and presumably alter the normal spatial regulation of abd-A products. We propose a model of abd-A structure based on a protein-coding region and two cis-regulatory regions. Regulatory region 1, 3′ to the transcription unit, contains positive and negative regulatory elements. Regulatory region 2, 5′ to the transcription unit, establishes the correct level of abd-A activity in the abdominal metameres.

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The organization and expression of the light gene, a heterochromatic gene of Drosophila melanogaster.

Genetics ◽

10.1093/genetics/125.1.129 ◽

1990 ◽

Vol 125 (1) ◽

pp. 129-140 ◽

Cited By ~ 6

Author(s):

R H Devlin ◽

B Bingham ◽

B T Wakimoto

Keyword(s):

Drosophila Melanogaster ◽

Dna Sequences ◽

Molecular Mapping ◽

Single Copy ◽

Transcription Unit ◽

Malpighian Tubules ◽

Centromeric Heterochromatin ◽

Chromosome 2 ◽

Induced Mutations ◽

Flanking Regions

Abstract The light (lt) gene is located in the centromeric heterochromatin of chromosome 2 of Drosophila melanogaster. This gene is necessary for normal levels of pigmentation in a number of adult and larval tissues and is required for viability. Hybrid dysgenic and X-ray induced mutations have been used to identify the gene and compare its organization to that of euchromatic genes. Molecular mapping of lt mutations and its major transcripts has shown that the lt gene is at least 17 kb. By injecting cosmid clones that include this region into lt mutant embryos, we have defined a 30-kb region that can transiently rescue the pigmentation defect in the Malpighian tubules. The major transcription unit of this gene is comprised of exons that are single copy. It is unusual in its organization in having a heterogeneous array of middle repetitive DNA sequences within its intronic and flanking regions.

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Molecular cloning and genetic analysis of the suppressor-of-white-apricot locus from Drosophila melanogaster

Molecular and Cellular Biology ◽

10.1128/mcb.7.7.2498-2505.1987 ◽

1987 ◽

Vol 7 (7) ◽

pp. 2498-2505

Author(s):

Z Zachar ◽

D Garza ◽

T B Chou ◽

J Goland ◽

P M Bingham

Keyword(s):

Drosophila Melanogaster ◽

Gene Transfer ◽

Genetic Analysis ◽

Developmental Stages ◽

Chromosomal Region ◽

Transcription Unit ◽

Complementation Group ◽

P Element ◽

Allele Specific ◽

New Mutations

We report genetic and molecular analysis of the suppressor-of-white-apricot [su(wa)] locus, one of several retrotransposon insertion allele-specific suppressor loci in Drosophila melanogaster. First, we isolated and characterized eight new mutations allelic to the original su(wa)1 mutation. These studies demonstrated that su(wa) mutations allelic to su(wa)1 affected a conventional D. melanogaster complementation group. Second, we cloned the chromosomal region containing the su(wa) complementation group by P element transposon tagging. The ca. 14-kilobase region surrounding the su(wa) complementation group contained five distinct transcription units, each with a different developmentally programmed pattern of expression. Third, we used a modified procedure for P-mediated gene transfer to identify the transcription unit corresponding to su(wa) by gene transfer. Fourth, we found that the presumptive su(wa) transcription unit produced a family of transcripts (ranging from ca. 3.5 to ca. 5.2 kilobases) in all developmental stages, tissue fractions, and cell lines we examined, suggesting that the gene is universally expressed.

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The complete sequence of the mouse skeletal alpha-actin gene reveals several conserved and inverted repeat sequences outside of the protein-coding region.

Molecular and Cellular Biology ◽

10.1128/mcb.6.1.15 ◽

1986 ◽

Vol 6 (1) ◽

pp. 15-25 ◽

Cited By ~ 52

Author(s):

M C Hu ◽

S B Sharp ◽

N Davidson

Keyword(s):

Amino Acids ◽

Nucleotide Sequence ◽

Inverted Repeat ◽

Actin Gene ◽

Coding Region ◽

Specific Expression ◽

Protein Coding ◽

Repeat Sequences ◽

Actin Genes ◽

Alpha Actin

The complete nucleotide sequence of a genomic clone encoding the mouse skeletal alpha-actin gene has been determined. This single-copy gene codes for a protein identical in primary sequence to the rabbit skeletal alpha-actin. It has a large intron in the 5'-untranslated region 12 nucleotides upstream from the initiator ATG and five small introns in the coding region at codons specifying amino acids 41/42, 150, 204, 267, and 327/328. These intron positions are identical to those for the corresponding genes of chickens and rats. Similar to other skeletal alpha-actin genes, the nucleotide sequence codes for two amino acids, Met-Cys, preceding the known N-terminal Asp of the mature protein. Comparison of the nucleotide sequences of rat, mouse, chicken, and human skeletal muscle alpha-actin genes reveals conserved sequences (some not previously noted) outside of the protein-coding region. Furthermore, several inverted repeat sequences, partially within these conserved regions, have been identified. These sequences are not present in the vertebrate cytoskeletal beta-actin genes. The strong conservation of the inverted repeat sequences suggests that they may have a role in the tissue-specific expression of skeletal alpha-actin genes.

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Molecular genetics of the Drosophila melanogaster ovo locus, a gene required for sex determination of germline cells.

Genetics ◽

10.1093/genetics/130.4.791 ◽

1992 ◽

Vol 130 (4) ◽

pp. 791-803

Author(s):

M D Garfinkel ◽

A R Lohe ◽

A P Mahowald

Keyword(s):

Drosophila Melanogaster ◽

Sex Determination ◽

Genomic Dna ◽

Transcription Unit ◽

Complementation Group ◽

P Element ◽

Female Sterility ◽

Germline Cells ◽

Female Specific ◽

Female Germline

Abstract The Drosophila melanogaster ovo gene is required for survival and differentiation of female germline cells, apparently playing a role in germline sex determination. We recovered 60 kb of genomic DNA from its genetic location at 4E1,2 on the X chromosome. A transcription unit coding for an apparently female-specific germline-dependent 5-kb poly(A)+ RNA size class is located substantially in a 7-kb region, within which three DNA-detectable lesions for mutations that inactivate the ovo function are located at two sites approximately 4 kb apart. The breakpoint of a deficiency that removes the neighboring lethal complementation group shavenbaby (svb) but leaves the ovo function intact maps approximately 5 kb to the molecular left of the leftmost ovo mutant site. A class of mutations that inactivates both the svb function and the ovo function affects genomic DNA between the two ovo sites. Sequences required for the two genetic functions are partly overlapping. In spite of this overlap, P element-mediated gene transfer of a 10-kb genomic DNA segment containing the 5-kb poly(A)+ RNA transcription unit rescues the female sterility phenotypes of ovo mutations, but not the svb lethality.

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SPONTANEOUS AND ETHYL METHANESULFONATE-INDUCED MUTATIONS CONTROLLING VIABILITY IN DROSOPHILA MELANOGASTER. I. RECESSIVE LETHAL MUTATIONS

Genetics ◽

10.1093/genetics/87.3.519 ◽

1977 ◽

Vol 87 (3) ◽

pp. 519-527

Author(s):

Ohmi Ohnishi

Keyword(s):

Drosophila Melanogaster ◽

Lethal Effect ◽

Induced Mutations ◽

Recessive Lethal ◽

Feeding Method ◽

Lethal Mutations ◽

Adult Feeding ◽

Lethal Rate ◽

Consistent Manner ◽

Mature Spermatozoa

ABSTRACT The efficiency of the adult feeding method for EMS treatment in Drosophila melanogaster was studied by measuring the frequency of induced recessive lethals on the second chromosome. The treatment was most effective when mature spermatozoa or spermatids were treated and was much less effective on earlier stages. The number of mutations induced was proportional to the concentration except at the highest doses. The recessive lethal rate was estimated to be about 0.012 per second chromosome per 10-4m. In addition, about 0.004-0.005 recessive lethals per 10-4 m were found in a later generation in chromosomes that had not shown the lethal effect in the previous generation. When the experiments are done in a consistent manner and gametes treated as mature sperm or spermatids are sampled, the results are highly reproducible. However, modifications of the procedure, such as starvation before EMS treatment, can considerably alter the effectiveness of the mutagen.

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