Abstract
Sildenafil improves erectile function by inhibiting the cGMP-catalytic activity of phosphodiesterase type V (PDE5). We used rapid amplification of cDNA Ends-polymerase chain reaction (RACE-PCR) to isolate three PDE5 isoforms from human corpus cavernosum. Semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis on eight human cavernous tissue samples showed that all samples expressed the PDE5A1 at a lower level than the PDE5A2 isoform. Five samples expressed the PDE5A3 isoform at various levels while the other three did not. Analysis on non-penile tissues showed that all tissues expressed the A1 and A2 isoforms while only those that have substantial amounts of smooth muscle expressed the A3 isoform. Cloning and sequencing of the PDE5A gene showed that the isoform-specific 5′-ends of the PDE5 mRNAs are encoded from three alternative first exons arranged in the order of A1–A3–A2. Promoter activities were detected upstream from the A1-specific exon and in the intron preceding the A2-specific exon. The upstream PDE5A promoter is expected to direct the expression of all three PDE5 isoforms while the intronic PDE5A2 promoter only the A2 isoform. Both promoters were upregulated by increasing concentrations of either cAMP or cGMP. Several transcription factor AP2 and Sp1-binding sequences identified in the promoters are likely to be the mediators of cAMP/cGMP-responsiveness.
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References
Schudt C, Gantner F, Tenors H, Hatzelmann A. . Therapeutic potential of selective PDE inhibitors in asthma. Pulm Pharmacol Ther 1999 12: 123–129.
Movsesian MA. . Beta-adrenergic receptor agonists and cyclic nucleotide phosphodiesterase inhibitors: shifting the focus from inotropy to cyclic adenosine monophosphate. J Am Coll Cardiol 1999 34: 318–324.
Ballard SA et al . Effects of sildenafil on the relaxation of human corpus cavernosum tissue in vitro and on the activities of cyclic nucleotide phosphodiesterase isozymes. J Urol 1998 159: 2164–2171.
Corbin JD, Francis SH. . Cyclic GMP phosphodiesterase-5: target of sildenafil. J Biol Chem 1999 274: 13729–13732.
Boolell M et al . Sildenafil: an orally active type 5 cyclic GMP-specific phosphodiesterase inhibitor for the treatment of penile erectile dysfunction. Int J Impot Res 1996 8: 47–52.
Francis SH, Corbin JD. . Purification of cGMP-binding protein phosphodiesterase from rat lung. Methods Enzymol 1988 159: 722–729.
Thomas MK, Francis SH, Corbin JD. . Characterization of a purified bovine lung cGMP-binding cGMP phosphodiesterase. J Biol Chem 1990 265: 14964–14970.
McAllister-Lucas LM et al . The structure of a bovine lung cGMP-binding, cGMP-specific phosphodiesterase deduced from a cDNA clone. J Biol Chem 1993 268: 22863–22873.
Kotera J et al . Expression of rat cGMP-binding cGMP-specific phosphodiesterase mRNA in Purkinje cell layers during postnatal neuronal development. Eur J Biochem 1997 249: 434–442.
Stacey P, Rulten S, Dapling A, Phillips SC. . Molecular cloning and expression of human cGMP-binding cGMP-specific phosphodiesterase (PDE5). Biochem Biophys Res Commun 1998 247: 249–254.
Loughney K et al . Isolation and characterization of cDNAs encoding PDE5A, a human cGMP-binding, cGMP-specific 3′, 5′-cyclic nucleotide phosphodiesterase. Gene 1998 216: 139–147.
Yanaka N et al . Expression, structure and chromosomal localization of the human cGMP-binding cGMP-specific phosphodiesterase PDE5A gene. Eur J Biochem 1998 255: 391–399.
Lin CS, Lau A, Tu R, Lue TF. . Expression of three isoforms of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in human penile cavernosum. Biochem Biophys Res Commun 2000 268: 628–635.
Lin CS, Lau A, Tu R, Lue TF. . Identification of three alternative first exons and an intronic promoter of human PDE5A gene. Biochem Biophys Res Commun 2000 268: 596–602.
Lin CS, Chang CH, Huynh T. . The murine L-plastin gene promoter: identification and comparison with the human L-plastin gene promoter. DNA Cell Biol 1997 16: 9–16.
Lin C-S, Aebersold RH, Leavitt J. . Correction of the N-terminal sequences of the human plastin isoforms by using anchored polymerase chain reaction: identification of a potential calcium-binding domain. Mol Cell Biol 1990 10: 1818–1821.
Thomson JM, Parrott WA. . pMECA: a cloning plasmid with 44 unique restriction sites that allows selection of recombinants based on colony size. BioTechniques 1998 24: 922–928.
Galas DJ, Schmitz A. . DNase footprinting: a simple method for the detection of protein-DNA binding specificity. Nucleic Acids Res 1978 5: 3157–3170.
Maxam AM, Gilbert W. . Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 1980 65: 499–560.
Kozak M. . The scanning model for translation: an update. J Cell Biol 1989 108: 229–241.
Stott K, Blackburn JM, Butler PJ, Perutz M. . Incorporation of glutamine repeats makes protein oligomerize: implications for neurodegenerative diseases. Proc Natl Acad Sci USA 1995 92: 6509–6513.
Nam RK et al . Significance of the CAG repeat polymorphism of the androgen receptor gene in prostate cancer progression. J Urol 2000 164: 567–572.
Sasaki M et al . The expansion of the CAG repeat in exon 1 of the human androgen receptor gene is associated with uterine endometrial carcinoma. Mol Carcinog 2000 27: 237–244.
Yu H et al . Shorter CAG repeat length in the androgen receptor gene is associated with more aggressive forms of breast cancer. Breast Cancer Res Treat 2000 59: 153–161.
Kawasaki T et al . Polymorphic CAG repeats of the androgen receptor gene and rheumatoid arthritis. Ann Rheum Dis 1999 58: 500–502.
Mifsud A et al . Trinucleotide (CAG) repeat polymorphisms in the androgen receptor gene: molecular markers of risk for male infertility. Fertil Steril 2001 75: 275–281.
Fischer KM. . Etiology of (CAG)n triplet repeat neurodegenerative diseases such as Huntington's disease is connected to stimulation of glutamate receptors. Med Hypotheses 1997 48: 393–398.
Bolger GB et al . Characterization of five different proteins produced by alternatively spliced mRNAs from the human cAMP-specific phosphodiesterase PDE4D gene. Biochem J 1997 328: 539–548.
Lin C-S et al . Age-related decrease of protein kinase G activation in vascular smooth muscle cells. Biochem Biophys Res Commun 2001 287: 244–248.
Vicini E, Conti M. . Characterization of an intronic promoter of a cyclic adenosine 3′, 5′-monophosphate (cAMP)-specific phosphodiesterase gene that confers hormone and cAMP inducibility. Mol Endocrinol 1997 11: 839–850.
Acknowledgements
This work was supported in part by grants from the California Urology Foundation (to C-S Lin) and from NIH (2R01-DK-45370, to TF Lue).
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Lin, CS., Chow, S., Lau, A. et al. Human PDE5A gene encodes three PDE5 isoforms from two alternate promoters. Int J Impot Res 14, 15–24 (2002). https://doi.org/10.1038/sj.ijir.3900802
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DOI: https://doi.org/10.1038/sj.ijir.3900802
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