Cyclic nucleotide PDEs (phosphodiesterases) are important enzymes that regulate intracellular levels of cAMP and cGMP. In the present study, we identify and characterize novel PDEs in the genetic model, Drosophila melanogaster. The Drosophila genome encodes five novel PDE genes in addition to dunce. Predicted PDE sequences of Drosophila show highly conserved critical domains when compared with human PDEs. Thus PDE-encoding genes of D. melanogaster are CG14940-PDE1C, CG8279-PDE6β, CG5411-PDE8A, CG32648-PDE9 and CG10231-PDE11. Reverse transcriptase–PCRs of adult tissues reveal widespread expression of PDE genes. Drosophila Malpighian (renal) tubules express all the six PDEs: Drosophila PDE1, dunce (PDE4), PDE6, PDE8, PDE9 and PDE11. Antipeptide antibodies were raised against PDE1, PDE6, PDE9 and PDE11. Verification of antibody specificity by Western blotting of cloned and expressed PDE constructs allowed the immunoprecipitation studies of adult Drosophila lysates. Biochemical characterization of immunoprecipitated endogenous PDEs showed that PDE1 is a dual-specificity PDE (Michaelis constant Km for cGMP: 15.3±1 μM; Km cAMP: 20.5±1.5 μM), PDE6 is a cGMP-specific PDE (Km cGMP: 37±13 μM) and PDE11 is a dual-specificity PDE (Km cGMP: 6±2 μM; Km cAMP: 18.5±5.5 μM). Drosophila PDE1, PDE6 and PDE11 display sensitivity to vertebrate PDE inhibitors, zaprinast (IC50 was 71±39 μM for PDE1, 0.65±0.015 μM for PDE6 and 1.6±0.5 μM for PDE11) and sildenafil (IC50 was 1.3±0.9 μM for PDE1, 0.025±0.005 μM for PDE6 and 0.12±0.06 μM for PDE11). We provide the first characterization of a cGMP-specific PDE and two dual-specificity PDEs in Drosophila, and show a high degree of similarity in structure and function between human and Drosophila PDEs.
Skip Nav Destination
Article navigation
May 2005
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
- PDF Icon PDF LinkEditorial Board
Research Article|
May 10 2005
Cyclic nucleotide phosphodiesterases in Drosophila melanogaster
Jonathan P. DAY;
Jonathan P. DAY
*Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 6NU, U.K.
Search for other works by this author on:
Julian A. T. DOW;
Julian A. T. DOW
*Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 6NU, U.K.
Search for other works by this author on:
Miles D. HOUSLAY;
Miles D. HOUSLAY
†Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 6NU, U.K.
Search for other works by this author on:
Shireen-A. DAVIES
Shireen-A. DAVIES
1
*Division of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 6NU, U.K.
1To whom correspondence should be addressed (email s.a.davies@bio.gla.ac.uk).
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
January 07 2005
Revision Received:
January 25 2005
Accepted:
January 27 2005
Accepted Manuscript online:
January 27 2005
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London
2005
Biochem J (2005) 388 (1): 333–342.
Article history
Received:
January 07 2005
Revision Received:
January 25 2005
Accepted:
January 27 2005
Accepted Manuscript online:
January 27 2005
Citation
Jonathan P. DAY, Julian A. T. DOW, Miles D. HOUSLAY, Shireen-A. DAVIES; Cyclic nucleotide phosphodiesterases in Drosophila melanogaster. Biochem J 15 May 2005; 388 (1): 333–342. doi: https://doi.org/10.1042/BJ20050057
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.