Originally published as Genetics Published Articles Ahead of Print on December 1, 2005.

Genetics, Vol. 172, 1621-1631, March 2006, Copyright © 2006
doi:10.1534/genetics.105.045641

The Purine Synthesis Gene Prat2 Is Required for Drosophila Metamorphosis, as Revealed by Inverted-Repeat-Mediated RNA Interference

Yingbiao Ji and Denise V. Clark1

Department of Biology, University of New Brunswick, Fredericton, New Brunswick E3B 5A3, Canada

1 Corresponding author: Department of Biology, University of New Brunswick, 10 Bailey Dr., Fredericton, NB E3B 5A3, Canada.
E-mail: clarkd{at}unb.ca

PRAT (phosphoribosylamidotransferase; E.C. 2.4.2.14) catalyzes the first reaction in de novo purine nucleotide biosynthesis. In Drosophila melanogaster, the Prat and Prat2 genes are both highly conserved with PRAT sequences from prokaryotes and eukaryotes. However, Prat2 organization and expression during development is different from Prat. We used RNA interference (RNAi) to knock down expression of both Prat and Prat2 to investigate their functions. Using the GAL4–UAS system, Prat RNAi driven by Act5c–GAL4 or tubP–GAL4 causes variable pupal lethality (48–100%) and ~50% female sterility, depending on the transgenic strains and drivers used. This observation agrees with the phenotype previously observed for Prat EMS-induced mutations. Prat2 RNAi driven by Act5C–GAL4 or tubP–GAL4 also results in variable pupal lethality (61–93%) with the different transgenic strains, showing that Prat2 is essential for fly development. However, Prat2 RNAi-induced arrested pupae have a head eversion defect reminiscent of the "cryptocephal" phenotype, whereas Prat RNAi-induced arrested pupae die later as pharate adults. We conclude that Prat2 is required during the prepupal stage while Prat is more important for the pupal stage. In addition, Prat and Prat2 double RNAi results in more severe pupal lethal phenotypes, suggesting that Prat and Prat2 have partially additive functions during Drosophila metamorphosis.