Uracil-containing DNA in Drosophila

Stability, stage-specific accumulation, and developmental involvement

Villo Muha, András Horváth, Angéla Békési, Mária Pukáncsik, Barbara Hodoscsek, Gábor Merényi, Gergely Róna, Júlia Batki, I. Kiss, Ferenc Jankovics, P. Vilmos, M. Erdélyi, B. Vértessy

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

Base-excision repair and control of nucleotide pools safe-guard against permanent uracil accumulation in DNA relying on two key enzymes: uracil-DNA glycosylase and dUTPase. Lack of the major uracil-DNA glycosylase UNG gene from the fruit fly genome and dUTPase from fruit fly larvae prompted the hypotheses that i) uracil may accumulate in Drosophila genomic DNA where it may be well tolerated, and ii) this accumulation may affect development. Here we show that i) Drosophila melanogaster tolerates high levels of uracil in DNA; ii) such DNA is correctly interpreted in cell culture and embryo; and iii) under physiological spatio-temporal control, DNA from fruit fly larvae, pupae, and imago contain greatly elevated levels of uracil (200-2,000 uracil/million bases, quantified using a novel real-time PCR-based assay). Uracil is accumulated in genomic DNA of larval tissues during larval development, whereas DNA from imaginal tissues contains much less uracil. Upon pupation and metamorphosis, uracil content in DNA is significantly decreased. We propose that the observed developmental pattern of uracil-DNA is due to the lack of the key repair enzyme UNG from the Drosophila genome together with down-regulation of dUTPase in larval tissues. In agreement, we show that dUTPase silencing increases the uracil content in DNA of imaginal tissues and induces strong lethality at the early pupal stages, indicating that tolerance of highly uracil-substituted DNA is also stage-specific. Silencing of dUTPase perturbs the physiological pattern of uracil-DNA accumulation in Drosophila and leads to a strongly lethal phenotype in early pupal stages. These findings suggest a novel role of uracil-containing DNA in Drosophila development and metamorphosis and present a novel example for developmental effects of dUTPase silencing in multicellular eukaryotes. Importantly, we also show lack of the UNG gene in all available genomes of other Holometabola insects, indicating a potentially general tolerance and developmental role of uracil-DNA in this evolutionary clade.

Original languageEnglish
Article numbere1002738
JournalPLoS Genetics
Volume8
Issue number6
DOIs
Publication statusPublished - Jun 2012

Fingerprint

uracil
Uracil
Drosophila
DNA
fruit flies
Uracil-DNA Glycosidase
Diptera
Fruit
Genome
genome
fruit
metamorphosis
Larva
insect larvae
repair
genomics
tolerance
Pupa
enzyme
larva

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology
  • Ecology, Evolution, Behavior and Systematics
  • Cancer Research
  • Genetics(clinical)

Cite this

Uracil-containing DNA in Drosophila : Stability, stage-specific accumulation, and developmental involvement. / Muha, Villo; Horváth, András; Békési, Angéla; Pukáncsik, Mária; Hodoscsek, Barbara; Merényi, Gábor; Róna, Gergely; Batki, Júlia; Kiss, I.; Jankovics, Ferenc; Vilmos, P.; Erdélyi, M.; Vértessy, B.

In: PLoS Genetics, Vol. 8, No. 6, e1002738, 06.2012.

Research output: Contribution to journalArticle

Muha, V, Horváth, A, Békési, A, Pukáncsik, M, Hodoscsek, B, Merényi, G, Róna, G, Batki, J, Kiss, I, Jankovics, F, Vilmos, P, Erdélyi, M & Vértessy, B 2012, 'Uracil-containing DNA in Drosophila: Stability, stage-specific accumulation, and developmental involvement', PLoS Genetics, vol. 8, no. 6, e1002738. https://doi.org/10.1371/journal.pgen.1002738
Muha, Villo ; Horváth, András ; Békési, Angéla ; Pukáncsik, Mária ; Hodoscsek, Barbara ; Merényi, Gábor ; Róna, Gergely ; Batki, Júlia ; Kiss, I. ; Jankovics, Ferenc ; Vilmos, P. ; Erdélyi, M. ; Vértessy, B. / Uracil-containing DNA in Drosophila : Stability, stage-specific accumulation, and developmental involvement. In: PLoS Genetics. 2012 ; Vol. 8, No. 6.
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