Developmental regulation of dUTPase in Drosophila melanogaster.
dUTPase prevents uracil incorporation into DNA by strict regulation of the cellular dUTP:dTTP ratio. Lack of the enzyme initiates thymineless cell death, prompting studies on enzyme regulation. We investigated expression pattern and localization of Drosophila dUTPase. Similarly to human, two isoforms of the fly enzyme were identified at both mRNA and protein levels. During larval stages, a drastic decrease of dUTPase expression was demonstrated at the protein level. In contrast, dUTPase mRNAs display constitutive character throughout development. A putative nuclear localization signal was identified in one of the two isoforms. However, immunohistochemistry of ovaries and embryos did not show a clear correlation between the presence of this signal and subcellular localization of the protein, suggesting that the latter may be perturbed by additional factors. Results are in agreement with a multilevel regulation of dUTPase in the Drosophila proteome, possibly involving several interacting protein partners of the enzyme. Using independent approaches, the existence of such macromolecular partners was verified.
Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, H-1518 Budapest, Hungary.
Angéla
A
Békési
Imre
I
Zagyva
Eva
E
Hunyadi-Gulyás
Veronika
V
Pongrácz
Júlia
J
Kovári
Agnes O
AO
Nagy
Anna
A
Erdei
Katalin F
KF
Medzihradszky
Beáta G
BG
Vértessy
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002453:Cell Cycle; D016923:Cell Death; D002460:Cell Line; D004330:Drosophila; D004331:Drosophila melanogaster / Q000201:enzymology*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D013058:Mass Spectrometry; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D010053:Ovary / Q000378:metabolism; D011233:Precipitin Tests; D020033:Protein Isoforms; D020543:Proteome; D011755:Pyrophosphatases / Q000096:biosynthesis* / Q000737:chemistry*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000378:metabolism; D020349:Surface Plasmon Resonance; D013997:Time Factors
D020033:Protein Isoforms; D020543:Proteome; D012333:RNA, Messenger; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase