The Drosophila melanogaster actin 5C gene uses two transcription initiation sites and three polyadenylation sites to express multiple mRNA species.

Bond, Beverley J.; Davidson, Norman

doi:10.1128/mcb.6.6.2080

Cited by 101 publications

(63 citation statements)

References 58 publications

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“…Neither a deletion of the Sex-lethal (Sx17BO) (22) gene nor a dominant allele of the double-sex gene (dsxD) (23) that can only be spliced to the male-specific splice site was able to suppress the sex comb defect in males homozygous for the P-element insertion (data not shown). We (28). RNAs were from adult male flies.…”

Section: Resultsmentioning

confidence: 99%

Mutations in the small subunit of the Drosophila U2AF splicing factor cause lethality and developmental defects.

Rudner

Kanaar

Breger

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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The essential eukaryotic pre-mRNA splicing factor U2AF (U2 small nuclear ribonucleoprotein auxiliary factor) is required to specify the 3' splice site at an early step in spliceosome assembly. U2AF binds site-specifically to the intron polypyrimidine tract and recruits U2 small nuclear ribonucleoprotein to the branch site. Human U2AF (hU2AF) is a heterodimer composed of a large (hU2AF65) and small (hU2AF35) subunit. Although these proteins associate in a tight complex, the biochemical requirement for U2AF activity can be satisfied solely by the large subunit. The requirement for the small subunit in splicing has remained enigmatic. No biochemical activity has been found for hU2AF35 and it has been implicated in splicing only indirectly by its interaction with known splicing factors. In the absence of a biochemical assay, we have taken a genetic approach to investigate the function of the small subunit in the fruit fly Drosophila melanogaster. A cDNA clone encoding the small subunit of Drosophila U2AF (dU2AF38) has been isolated and sequenced. The dU2AF38 protein is highly homologous to hU2AF35 containing a conserved central arginine-and serine-rich (RS) domain. A recessive P-element insertion mutation affecting dU2AF38 causes a reduction in viability and fertility and morphological bristle defects. Consistent with a general role in splicing, a null allele of dU2AF38 is fully penetrant recessive lethal, like null alleles of the Drosophila U2AF large subunit.Generation of functional mRNA in eukaryotes requires the removal of noncoding regions (introns) from pre-mRNA by a process termed RNA splicing (1-3). Pre-mRNA splicing takes place in the spliceosome, a dynamic RNA-protein complex that assembles in a stepwise ATP-dependent manner on the pre-mRNA (1-3). The spliceosome is composed of small nuclear ribonucleoprotein particles (snRNPs) and extrinsic (non-snRNP) protein factors. Studies with human cell (HeLa) nuclear splicing extracts have shown that targeting of U2 snRNP to the branch site on the pre-mRNA, an early step in spliceosome assembly, requires a protein factor called U2AF (U2 snRNP auxiliary factor) (4). U2AF binds site-specifically to the intron polypyrimidine tract located between the branch site and the 3' splice site of the pre-mRNA (5). U2AF recruits U2 snRNP to the branch site in the first ATP-dependent step in spliceosome assembly. Recruitment of U2 snRNP to the branch site defines the 3' splice site since in most cases the first AG dinucleotide downstream of the branch site is used as the 3' splice site (6). Thus, U2AF plays a critical role in 3' splice site selection.Human U2AF (hU2AF) consists of two polypeptides, a 65-kDa large subunit (hU2AF65) and a 35-kDa small subunit (hU2AF35) (7). hU2AF65 contains three carboxyl-terminal ribonucleoprotein consensus sequence (RNP-CS) domains that mediate RNA binding and an amino-terminal arginineand serine-rich (RS) domain (5). All domains are essential for splicing in vitro (5). hU2AF35 contains a central RS domain and a carboxyl-terminal glycine...

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Section: Resultsmentioning

confidence: 99%

Mutations in the small subunit of the Drosophila U2AF splicing factor cause lethality and developmental defects.

Rudner

Kanaar

Breger

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

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“…5. The lane containing ovary RNA was exposed for 2 h, the lane containing testis RNA was exposed for 24 h, and the lanes containing RNA from heads and thoraces were exposed for 7 …”

Section: Resultsmentioning

confidence: 99%

Molecular localization and developmental expression of the otu locus of Drosophila melanogaster.

Mulligan¹,

Mohler²,

Kalfayan³

1988

Mol. Cell. Biol.

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The female-sterile ovarian tumor gene, otu, is located in cytological region 7F1 on the Drosophila melanogaster chromosome map. We have mapped the gene at the molecular level by using four dysgenic alleles and two revertant derivatives of these alleles as well as an ethyl methanesulfonate-induced allele. The insertional (dysgenic) changes were all associated with one restriction fragment, and its size was restored after phenotypic reversion. One ethyl methanesulfonate-induced allele had a deletion in the restriction fragment adjacent (distal) to the fragment altered in the insertional alleles. These two restriction fragments were immediately adjacent to the s38 chorion gene. Associated with the two altered restriction fragments were two RNA species, an abundant 3.2-kilobase (kb) poly(A)+ RNA and a minor 4.0-kb RNA. Several other less-abundant RNA species were detectable with more-sensitive single-stranded RNA probes. The otu gene was transcribed proximal to distal relative to the centromere; this was opposite to the direction of transcription of the adjacent s38 gene. During development, the 3.2-kb RNA was absent in larvae, first appeared in the pupal stages, and persisted in adult females, in which it was most prevalent in the ovaries. The DNA that hybridized to the 3.2-kb ovarian RNA hybridized to four different RNAs found in the testes but not in the rest of the adult male. These testis-enriched RNAs were transcribed from the same strand of DNA as the ovarian transcripts.Our understanding of germ cell determination and differentiation in Drosophila melanogaster oogenesis is derived largely from morphological and genetic studies. During oogenesis, a single apical ovarian cell, the germarial cystoblast, undergoes four incomplete cell divisions to produce a syncytium of 16 interconnected cystocytes. One of these cystocytes differentiates into an oocyte (19), while the remaining 15 become the nurse cells that nourish the growing oocyte via intercellular cytoplasmic bridges (8,(26)(27)(28) and provide it with RNAs and proteins that will be required during early embryogenesis (30).Of central interest in germ cell development is the mechanism by which 1 cystocyte develops into an oocyte and the other 15 cystocytes develop into nurse cells. Within a 16-cell syncytium, the number of intercellular bridges varies from one to four per cell, with only 2 of the 16 cystocytes having four bridges. Immediately after the formation of the 16-cell cluster, both four-canal cells enter meiotic prophase (27, 28). However, one of these cells subsequently enters the nurse cell developmental pathway, while the other develops as an oocyte. It therefore appears that four cytoplasmic bridges are a necessary but not sufficient prerequisite for determination of oocyte development.We are interested in the early events of oogenesis and specifically in genes that regulate the morphogenesis of the germ cells themselves. The otu gene is one of several genes whose products are required for the determination and differentiation of oocytes and nur...

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“…(A similar approach was devised independently by Ft. Holmgren [personal communication].) In brief, a constitutive promoter, AcfSC (Actin in the 5C region; Bond and Davidson, 1986), is placed upstream of a given coding sequence (/acZ or wg) but separated from it by a segment of DNA bounded by direct FRT repeats that we call a f/p-out cassette. If, as diagramed, the intervening DNA contains a transcriptional stop, the coding sequence will not be expressed unless the cassette is excised by f/p-mediated recombination.…”

Section: A General Methods For Heritably Activating Any Gene In Any Cellmentioning

confidence: 99%

“…Act% A Noti linker was inserted at the EcoRl site positioned -4.4 kb upstream of the translational start of the Act% gene (Bond and Davidson, 1986). A set of oiigonucieotides kindly provided by 8.…”

Section: Experlmental Procedures Piasmid Constructionsmentioning

confidence: 99%

Organizing activity of wingless protein in Drosophila

Struhl

Basler

1993

Cell

816

566

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The adult appendages of Drosophila are formed from imaginal discs, sheets of epithelial cells that proliferate during larval development and differentiate during metamorphosis. wingless (wg, DWnt-1) protein, a putative signaling molecule, is expressed only in prospective ventral cells in each of the leg discs. To test the role of wg, we have generated randomly positioned clones of cells that express wg protein constitutively. Clones that arise in the prospective ventral portions of the leg discs develop normally. In contrast, dorsally situated clones give rise to ventrolateral patterns and exert a ventralizing influence on neighboring wild-type tissue. We propose that wg protein organizes leg pattern along the dorsoventral axis by conferring ventral positional information within the disc. The adult appendages of Drosophila are formed from imaginal discs, sheets of epithelial cells that proliferate during larval development and differentiate during metamorphosis. wingless (wg, DWnt-1) protein, a putative signaling molecule, is expressed only in prospective ventral cells in each of the leg discs. To test the role of wg, we have generated randomly positioned clones of cells that express wg protein constitutively. Clones that arise in the prospective ventral portions of the leg discs develop normally. In contrast, dorsally situated clones give rise to ventrolateral patterns and exert a ventraliring influence on neighboring wild-type tissue. We propose that wg protein organizes leg pattern along the dorsoventral axis by conferring ventral positional information within the disc.

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The Drosophila melanogaster actin 5C gene uses two transcription initiation sites and three polyadenylation sites to express multiple mRNA species.

Cited by 101 publications

References 58 publications

Mutations in the small subunit of the Drosophila U2AF splicing factor cause lethality and developmental defects.

Mutations in the small subunit of the Drosophila U2AF splicing factor cause lethality and developmental defects.

Molecular localization and developmental expression of the otu locus of Drosophila melanogaster.

Organizing activity of wingless protein in Drosophila

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