Regulation of a Drosophila melanogaster cGMP-specific phosphodiesterase by prenylation and interaction with a prenyl-binding protein

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)

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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
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 > Institute of Molecular Cell and Systems Biology
College of Medical Veterinary and Life Sciences > Institute of Neuroscience and Psychology
Journal Name:Biochemical Journal
Publisher:Portland Press Ltd.
ISSN:0264-6021
ISSN (Online):1470-8728
Published Online:27 May 2008
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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
438301Phosphodiesterase-4 isoforms - intracellular targeting, regulation and potential therapeutic targetsMiles HouslayMedical Research Council (MRC)G0600765Institute of Neuroscience and Psychology
378481Exploration of cyclic 3', 5' guanosine monophosphate (cGMP) signalling dynamics in vivoShireen DaviesBiotechnology and Biological Sciences Research Council (BBSRC)BB/C000633/1Institute of Molecular Cell and Systems Biology