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Abstract
The pugilist-Dominant mutation results from fusion of a portion of the gene encoding the tri-functional Methylene Tetrahydrofolate Dehydrogenase (E.C.1.5.1.5, E.C.3.5.4.9, E.C.6.3.4.3) to approximately one kb of a heterochromatic satellite repeat. Expression of this fusion gene results in an unusual ring pattern of pigmentation around the eye. We carried out experiments to determine the mechanism for this pattern. By using FLP-mediated DNA mobilization to place different pugD transgenes at pre-selected sites we found that variation in repeat length makes a strong contribution to variability of the pug phenotype. This variation is manifest primarily as differences in the thickness of the pigmented ring. We show that similar phenotypic variation can also be achieved by changing gene copy number. We found that the pugD pattern is not controlled by wingless, which is normally expressed in a similar ring pattern. Finally, we found that physical injury to a pugD eye can lead to pigment deposition in parts of the eye that would not have been pigmented in the absence of injury. Our results are consistent with a model in which a metabolite vital for pigment formation is imported from the periphery of the eye, and pugD limits the extent of its transport towards the center of the eye, thus revealing the existence of a hitherto unknown mechanism of localized transport in the eye.
Highlights
The pugilist gene of Drosophila melanogaster encodes an enzyme with three activities in tetrahydrofolate metabolism
Independent insertions of a pugD transgene carried within a P element produce varied eye pigment phenotypes, differing primarily in the diameter of the central region that lacks pteridines
We previously showed that a pug transgene with only 300 bp of AGAGAGA repeats was ineffective at producing the pugD phenotype [1].To test this, we made use of P element doi:10.1371/journal.pone.0151377.g002
Summary
The pugilist (pug) gene of Drosophila melanogaster encodes an enzyme with three activities in tetrahydrofolate metabolism. The dominant pugD mutation reduces pigment throughout the eye, with an effect that is especially noticeable in a background where only pteridines are present, e.g. vermillion (v). The eyes of such flies (v; pugD/+) exhibit an unusual ring of pigmentation around the periphery of the eye and a few scattered spots of pigment in the center of the eye, but are otherwise completely white-eyed. The protein encoded by pug has been implicated in the response to Parkinson's disease, aging and oxidative stress, immunity [2,3,4,5,6]
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