Biomedical subjects

Kalpana White

Publications and source records attributed to Kalpana White.

6 recordsLinked to original sources

PubMed2006

Sex-peptide-regulated female sexual behavior requires a subset of ascending ventral nerve cord neurons.

BACKGROUND: Male-derived Sex-peptide (SP) elicits egg laying and rejection of courting males in mated Drosophila females. Little is known about the genes that specify the underlying neuronal circuits and mediate this switch in female sexual behavior. RESULTS: Here we show that the egghead gene involved in glycosphingolipid biosynthesis provides an essential component to the SP response. We have isolated viable alleles of the vital egghead gene that abolish egghead expression from a distal promoter resulting in the absence of the largest transcript of this complex transcription unit. Temporally and spatially restricted expression of egghead revealed a requirement for egghead early in the development of apterous-expressing ventral nerve cord neurons to rescue the SP response. In viable egghead alleles, these ascending interneurons, three per abdominal and seven per thoracic hemisegment, fail to innervate the central brain. egghead expression in apterous neurons rescues neuronal targeting and the response to SP. Furthermore, neurotransmission in apterous neurons is required to elicit the SP response. CONCLUSION: Together with the former finding of SP binding to afferent nerves , these results suggest that SP-mediated modification of sensory input switches female sexual behavior from the virgin to the mated state.

Amino Acid Sequence↗

PubMed2005

The egghead gene is required for compartmentalization in Drosophila optic lobe development.

The correct targeting of photoreceptor neurons (R-cells) in the developing Drosophila visual system requires multiple guidance systems in the eye-brain complex as well as the precise organization of the target area. Here, we report that the egghead (egh) gene, encoding a glycosyltransferase, is required for a compartment boundary between lamina glia and lobula cortex, which is necessary for appropriate R1-R6 innervation of the lamina. In the absence of egh, R1-R6 axons form a disorganized lamina plexus and some R1-R6 axons project abnormally to the medulla instead of the lamina. Mosaic analysis demonstrates that this is not due to a loss of egh function in the eye or in the neurons and glia of the lamina. Rather, as indicated by clonal analysis and cell-specific genetic rescue experiments, egh is required in cells of the lobula complex primordium which transiently abuts the lamina and medulla in the developing larval brain. In the absence of egh, perturbation of sheath-like glial processes occurs at the boundary region delimiting lamina glia and lobula cortex, and inappropriate invasion of lobula cortex cells across this boundary region disrupts the pattern of lamina glia resulting in inappropriate R1-R6 innervation. This finding underscores the importance of the lamina/lobula compartment boundary in R1-R6 axon targeting.

Animals↗

PubMed2005

ELAV multimerizes on conserved AU4-6 motifs important for ewg splicing regulation.

ELAV is a gene-specific regulator of alternative pre-mRNA processing in Drosophila neurons. Since ELAV/Hu proteins preferentially bind to AU-rich regions that are generally abundant in introns and untranslated regions, it has not been clear how gene specificity is achieved. Here we used a combination of in vitro biochemical experiments together with phylogenetic comparisons and in vivo analysis of Drosophila transgenes to study ELAV binding to the last ewg intron and splicing regulation. In vitro binding studies of ELAV show that ELAV multimerizes on the ewg binding site and forms a defined and saturable complex. Further, sizing of the ELAV-RNA complex and a series of titration experiments indicate that ELAV forms a dodecameric complex on 135 nucleotides in the last ewg intron. Analysis of the substrate RNA requirements for ELAV binding and complex formation indicates that a series of AU(4-6) motifs spread over the entire binding site are important, but not a strictly defined sequence element. The importance of AU(4-6) motifs, but not spacing between them, is further supported by evolutionary conservation in several melanogaster species subgroups. Finally, using transgenes we demonstrate in fly neurons that ELAV-mediated regulation of ewg intron 6 splicing requires several AU(4-6) motifs and that introduction of spacer sequence between conserved AU(4-6) motifs has a minimal effect on splicing. Collectively, our results suggest that ELAV multimerization and binding to multiple AU(4-6) motifs contribute to target RNA recognition and processing in a complex cellular environment.

AT Rich Sequence↗

PubMed2004

ELAV.

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Alternative Splicing↗

PubMed2003

ELAV inhibits 3'-end processing to promote neural splicing of ewg pre-mRNA.

The embryonic lethal abnormal visual system (ELAV) is a gene-specific regulator of alternative pre-mRNA processing in neurons of Drosophila. Here we define a functional in vivo binding site for ELAV in neurons through the development of a reporter gene system in transgenic animals in combination with in vitro binding assays. ELAV binds to erect wing (ewg) RNA 3' of a polyadenylation site in the terminal intron 6. At this polyadenylation site, ELAV inhibits 3'-end processing in vitro in a dose-dependent and sequence-specific manner, and ELAV binding is necessary in vivo to promote splicing of ewg intron 6. Further, the AAUAAA poly(A) complex recognition sequence, together with ELAV, is required to regulate neural 3' splice site choice in vivo. In addition, the use of segmentally labeled RNA substrates in UV cross-linking assays suggest that ELAV does not inhibit or redirect binding of cleavage factor dCstF64 at the regulated polyadenylation site on ewg RNA. These data indicate that binding of 3'-end processing factors, together with ELAV, can regulate alternative splicing.

Adenosine↗

PubMed2002

Ectopic expression of Drosophila ELAV and human HuD in Drosophila wing disc cells reveals functional distinctions and similarities.

Drosophila ELAV and human HuD are two neuronal RNA binding proteins that show remarkable sequence homology, yet differ in their respective documented roles in post-transcriptional regulation. ELAV regulates neural-specific alternative splicing of specific transcripts, and HuD stabilizes specific mRNAs that are otherwise unstable due to AU-rich elements (AREs) in their 3' untranslated region (UTR). AREs are major determinants of transcript stability in mammalian cells. The role of each of these proteins was investigated and compared, by ectopically expressing them in Drosophila imaginal wing disc cells, which lack endogenous expression of either protein. The effect of the ectopic expression of ELAV and HuD was assessed on two sets of green fluorescent protein reporter transgenes, which were all driven with a broadly expressing promoter. Each set consisted of three reporter transgenes: (1) with an uninterrupted open reading frame (ORF); (2) with a constitutively spliced intron inserted into the ORF; and (3) with the intron nASI whose splicing is regulated in neurons by ELAV, inserted into the ORF. The two sets differed from each other only in their 3'UTR: Heat-shock-protein-70Ab (Hsp70Ab) trailer with ARE-like characteristics or Actin 5C (Act5C) trailer. Our results show that: (1) both ectopically expressed ELAV and HuD can enhance expression of transgenes with the Hsp70Ab 3'UTR, but not of transgenes with Act5C 3'UTR; (2) this enhancement is accompanied by an increase in mRNA level; (3) only ELAV can induce neural-specific splicing of nASI; and (4) although HuD is localized primarily to the cytoplasm, ELAV is localized to both the cytoplasm and the nucleus.

3' Untranslated Regions↗