A P-Element Insertion Screen Identified Mutations in 455 Novel Essential Genes in Drosophila

Abstract

With the completion of the nucleotide sequences of several complex eukaryotic genomes, tens of thousands of genes have been predicted. However, this information has to be correlated with the functions of those genes to enhance our understanding of biology and to improve human health care. The Drosophila transposon P-element-induced mutations are very useful for directly connecting gene products to their biological function. We designed an efficient transposon P-element-mediated gene disruption procedure and performed genetic screening for single P-element insertion mutations, enabling us to recover 2500 lethal mutations. Among these, 2355 are second chromosome mutations. Sequences flanking >2300 insertions that identify 850 different genes or ESTs (783 genes on the second chromosome and 67 genes on the third chromosome) have been determined. Among these, 455 correspond to genes for which no lethal mutation has yet been reported. The Drosophila genome is thought to contain ∼3600 vital genes; 1400 are localized on the second chromosome. Our mutation collection represents ∼56% of the second chromosome vital genes and ∼24% of the total vital Drosophila genes.

Allele-specific alternative splicing of Drosophila Ribosomal protein S21 suppresses a lethal mutation in the Phosphorylated adaptor for RNA export (Phax) gene

“…1, a and b ). Prior to our rescue experiments and the site-specific integration of the UAS - Phax transgene into the Phax SH/SH mutant background, adult escapers, homozygous for the P element insertion in the Phax gene, were intermittently detectable in different stocks of this Phax SH/SH mutant line ( Oh et al 2003 ; Kahsai et al 2016 ; Garcia et al 2016 ). We hypothesized that subtle genetic background differences might contribute to the observed differences in the viability of different stocks of the Phax SH/SH mutant and in our transgenic rescue experiments.…”

“…Harrison. Recombination of the UAS:Phax-mVenus transgene into the mutant (P{lacW} Phax SH0641 ) ( Oh et al 2003 ) was previously described ( Garcia et al 2016 ). Here, Phax SH/SH refers to homozygous lines with the P element insertion in the Phax gene (P{lacW} Phax SH0641 ).…”

“…Full loss of fly Smn function results in a recessive larval lethality in Drosophila ( Chan et al 2003 ; Rajendra et al 2007 ; Chang et al 2008 ). In contrast, lethality can be genetically separated from a disruptive P element insertion in the Drosophila Phax gene, an insertion previously classified as lethal ( Oh et al 2003 ; Kahsai et al 2016 ). Our lab and others have observed adult flies that are homozygous for this P element insertion in the Phax gene ( Kahsai et al 2016 ).…”

The ubiquitin thioesterase YOD1 ameliorates mutant Huntingtin induced pathology in Drosophila

“…Generation of Usp5 2 deletion mutants were described in 42 . Deletion null alleles for Yod1 and Duba were generated by standard P-element remobilization technique 43 . In brief, flies carrying the P-element were crossed to flies expressing the P{Δ2-3} transposase.…”

Du13 encodes a C2H2 zinc‐finger protein that regulates Wxb pre‐mRNA splicing and microRNA biogenesis in rice endosperm

“…Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd., 20, 1387-1401 served a basic cellular function. Null alleles of SE homologs confer embryonic lethality in Arabidopsis, yeast (Schizosaccharomyces pombe), fruit fly (Drosophila melanogaster), zebrafish (Danio rerio) and mouse (Mus musculus; Amsterdam et al, 2004;Golling et al, 2002;Kim et al, 2010;Lobbes et al, 2006;Oh et al, 2003;Wilson et al, 2008). Most eukaryotic genomes encode a single SE homolog, whereas monocot genomes harbour multiple SE homologs (Figure S5a).…”