Day, J. P., Cleghon, V., Houslay, M. D. and Davies, S. A. (2008) Regulation of a Drosophila melanogaster cGMP-specific phosphodiesterase by prenylation and interaction with a prenyl-binding protein. Biochemical Journal, 414(Pt3), pp. 363-374. (doi: 10.1042/BJ20080560) (PMID:18503409)
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Publisher's URL: http://dx.doi.org/10.1042/BJ20080560
Abstract
Post-translational modification by isoprenylation is a pivotal process for the correct functioning of many signalling proteins. The <i>Drosophila</i> <i>melanogaster</i> cGMP-PDE (cGMP-specific phosphodiesterase) <i>Dm</i>PDE5/6 possesses a C<i>aa</i>X-box prenylation signal motif, as do several novel cGMP-PDEs from insect and echinoid species (in C<i>aa</i>X, C is cysteine, a is an aliphatic amino acid and X is 'any' amino acid). D<i>m</i>PDE5/6 is prenylated <i>in vivo</i> at Cys(1128) and is localized to the plasma membrane when expressed in <i>Drosophila</i> S2 cells. Site-directed mutagenesis of the prenylated cysteine residue (C1128S-DmPDE5/6), pharmacological inhibition of prenylation or co-expression of D<i>m</i>PrBP (Drosophila prenyl-binding protein)/δ each alters the subcellular-localization of D<i>m</i>PDE5/6. Thus prenylation constitutes a critical post-translational modification of D<i>m</i>PDE5/6 for membrane targeting. Co-immunoprecipitation and subcellular-fractionation experiments have shown that D<i>m</i>PDE5/6 interacts with D<i>m</i>PrBP/δ in Drosophila S2 cells. Transgenic lines allow targeted expression of tagged prenylation-deficient C1128S-D<i>m</i>PDE5/6 in Type I (principal) cells in Drosophila Malpighian tubules, ail <i>in vivo</i> model for D<i>m</i>PDE5/6 function. In contrast with wild-type D<i>m</i>PDE5/6, which was exclusively associated with the apical membrane, the C1128S-D<i>m</i>PDE5/6 Mutant form was located primarily in the cytosol, although some residual association occurred at the apical membrane. Despite the profound change in intracellular localization of C1128S-D<i>m</i>PDE5/6, active transport of cGMP is affected in the same way as it is by D<i>m</i>PDE5/6. This suggests that, in addition to prenylation and interaction with D<i>m</i>PrBP/δ, further functional membrane-targeting signals exist within D<i>m</i>PDE5/6
Item Type: | Articles |
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Status: | Published |
Refereed: | Yes |
Glasgow Author(s) Enlighten ID: | Houslay, Professor Miles and Day, Dr Jonathan and Davies, Professor Shireen |
Authors: | Day, J. P., Cleghon, V., Houslay, M. D., and Davies, S. A. |
Subjects: | Q Science > QH Natural history > QH345 Biochemistry |
College/School: | College of Medical Veterinary and Life Sciences > School of Molecular Biosciences College of Medical Veterinary and Life Sciences > School of Psychology & Neuroscience |
Journal Name: | Biochemical Journal |
Publisher: | Portland Press Ltd. |
ISSN: | 0264-6021 |
ISSN (Online): | 1470-8728 |
Published Online: | 27 May 2008 |
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