Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 2002 Jan 15;361(Pt 2):211–220. doi: 10.1042/0264-6021:3610211

Novel alternative splice variants of rat phosphodiesterase 7B showing unique tissue-specific expression and phosphorylation.

Takashi Sasaki 1, Jun Kotera 1, Kenji Omori 1
PMCID: PMC1222301  PMID: 11772393

Abstract

cDNA species coding for novel variants of cyclic-AMP-specific phosphodiesterases (PDEs), namely the PDE7B family, were isolated from rats and characterized. Rat PDE7B1 (RNPDE7B1) was composed of 446 amino acid residues. Rat PDE7B2 (RNPDE7B2) and PDE7B3 (RNPDE7B3), which possessed unique N-terminal sequences, consisted of 359 and 459 residues respectively. Northern hybridization analysis showed that rat PDE7B transcripts were particularly abundant in the striatum and testis. PCR analyses revealed that rat PDE7B2 transcripts were restricted to the testis and that low levels of PDE7B3 transcripts were expressed in the heart, lung and skeletal muscle. In situ hybridization analysis demonstrated that rat PDE7B transcripts were expressed in striatal neurons and spermatocytes. In spermatocytes, rat PDE7B transcripts were expressed in a stage-specific manner during spermatogenesis. The K(m) values of recombinant rat PDE7B1, PDE7B2 and PDE7B3 for cAMP were 0.05, 0.07 and 0.05 microM respectively. Each rat PDE7B variant was the most sensitive to 3-isobutyl-1-methylxanthine (IC(50) 1.5-2.1 microM). Two phosphorylation sites for cAMP-dependent protein kinase (PKA) were found in rat PDE7B1 and PDE7B3, whereas rat PDE7B2 possessed one site. PKA-dependent phosphorylation was observed in C-terminal phosphorylation sites of three rat PDE7B variants, in addition to unique N-terminal regions of rat PDE7B1 and PDE7B3. Unique tissue distribution and PKA-dependent phosphorylation of PDE7B variants suggested that each variant has a specific role for cellular functions via cAMP signalling in various tissues.

Full Text

The Full Text of this article is available as a PDF (443.9 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Andrews T. C., Weeks R. A., Turjanski N., Gunn R. N., Watkins L. H., Sahakian B., Hodges J. R., Rosser A. E., Wood N. W., Brooks D. J. Huntington's disease progression. PET and clinical observations. Brain. 1999 Dec;122(Pt 12):2353–2363. doi: 10.1093/brain/122.12.2353. [DOI] [PubMed] [Google Scholar]
  2. Beavo J. A. Cyclic nucleotide phosphodiesterases: functional implications of multiple isoforms. Physiol Rev. 1995 Oct;75(4):725–748. doi: 10.1152/physrev.1995.75.4.725. [DOI] [PubMed] [Google Scholar]
  3. Bibb J. A., Yan Z., Svenningsson P., Snyder G. L., Pieribone V. A., Horiuchi A., Nairn A. C., Messer A., Greengard P. Severe deficiencies in dopamine signaling in presymptomatic Huntington's disease mice. Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6809–6814. doi: 10.1073/pnas.120166397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Burns F., Zhao A. Z., Beavo J. A. Cyclic nucleotide phosphodiesterases: gene complexity, regulation by phosphorylation, and physiological implications. Adv Pharmacol. 1996;36:29–48. doi: 10.1016/s1054-3589(08)60575-x. [DOI] [PubMed] [Google Scholar]
  5. Cha J. H., Kosinski C. M., Kerner J. A., Alsdorf S. A., Mangiarini L., Davies S. W., Penney J. B., Bates G. P., Young A. B. Altered brain neurotransmitter receptors in transgenic mice expressing a portion of an abnormal human huntington disease gene. Proc Natl Acad Sci U S A. 1998 May 26;95(11):6480–6485. doi: 10.1073/pnas.95.11.6480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Degerman E., Belfrage P., Manganiello V. C. Structure, localization, and regulation of cGMP-inhibited phosphodiesterase (PDE3). J Biol Chem. 1997 Mar 14;272(11):6823–6826. doi: 10.1074/jbc.272.11.6823. [DOI] [PubMed] [Google Scholar]
  7. Francis S. H., Turko I. V., Corbin J. D. Cyclic nucleotide phosphodiesterases: relating structure and function. Prog Nucleic Acid Res Mol Biol. 2001;65:1–52. doi: 10.1016/s0079-6603(00)65001-8. [DOI] [PubMed] [Google Scholar]
  8. Fujishige K., Kotera J., Michibata H., Yuasa K., Takebayashi S., Okumura K., Omori K. Cloning and characterization of a novel human phosphodiesterase that hydrolyzes both cAMP and cGMP (PDE10A). J Biol Chem. 1999 Jun 25;274(26):18438–18445. doi: 10.1074/jbc.274.26.18438. [DOI] [PubMed] [Google Scholar]
  9. Greengard P., Allen P. B., Nairn A. C. Beyond the dopamine receptor: the DARPP-32/protein phosphatase-1 cascade. Neuron. 1999 Jul;23(3):435–447. doi: 10.1016/s0896-6273(00)80798-9. [DOI] [PubMed] [Google Scholar]
  10. Han P., Zhu X., Michaeli T. Alternative splicing of the high affinity cAMP-specific phosphodiesterase (PDE7A) mRNA in human skeletal muscle and heart. J Biol Chem. 1997 Jun 27;272(26):16152–16157. doi: 10.1074/jbc.272.26.16152. [DOI] [PubMed] [Google Scholar]
  11. Harrison M. B., Wiley R. G., Wooten G. F. Changes in D2 but not D1 receptor binding in the striatum following a selective lesion of striatopallidal neurons. Brain Res. 1992 Sep 11;590(1-2):305–310. doi: 10.1016/0006-8993(92)91111-q. [DOI] [PubMed] [Google Scholar]
  12. Hemmings H. C., Jr, Greengard P., Tung H. Y., Cohen P. DARPP-32, a dopamine-regulated neuronal phosphoprotein, is a potent inhibitor of protein phosphatase-1. Nature. 1984 Aug 9;310(5977):503–505. doi: 10.1038/310503a0. [DOI] [PubMed] [Google Scholar]
  13. Hetman J. M., Soderling S. H., Glavas N. A., Beavo J. A. Cloning and characterization of PDE7B, a cAMP-specific phosphodiesterase. Proc Natl Acad Sci U S A. 2000 Jan 4;97(1):472–476. doi: 10.1073/pnas.97.1.472. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hoffmann R., Baillie G. S., MacKenzie S. J., Yarwood S. J., Houslay M. D. The MAP kinase ERK2 inhibits the cyclic AMP-specific phosphodiesterase HSPDE4D3 by phosphorylating it at Ser579. EMBO J. 1999 Feb 15;18(4):893–903. doi: 10.1093/emboj/18.4.893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hoffmann R., Wilkinson I. R., McCallum J. F., Engels P., Houslay M. D. cAMP-specific phosphodiesterase HSPDE4D3 mutants which mimic activation and changes in rolipram inhibition triggered by protein kinase A phosphorylation of Ser-54: generation of a molecular model. Biochem J. 1998 Jul 1;333(Pt 1):139–149. doi: 10.1042/bj3330139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huang S. H., Pittler S. J., Huang X., Oliveira L., Berson E. L., Dryja T. P. Autosomal recessive retinitis pigmentosa caused by mutations in the alpha subunit of rod cGMP phosphodiesterase. Nat Genet. 1995 Dec;11(4):468–471. doi: 10.1038/ng1295-468. [DOI] [PubMed] [Google Scholar]
  17. Jin S. L., Richard F. J., Kuo W. P., D'Ercole A. J., Conti M. Impaired growth and fertility of cAMP-specific phosphodiesterase PDE4D-deficient mice. Proc Natl Acad Sci U S A. 1999 Oct 12;96(21):11998–12003. doi: 10.1073/pnas.96.21.11998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kangasniemi M., Kaipia A., Mali P., Toppari J., Huhtaniemi I., Parvinen M. Modulation of basal and FSH-dependent cyclic AMP production in rat seminiferous tubules staged by an improved transillumination technique. Anat Rec. 1990 May;227(1):62–76. doi: 10.1002/ar.1092270108. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Le Moine C., Bloch B. D1 and D2 dopamine receptor gene expression in the rat striatum: sensitive cRNA probes demonstrate prominent segregation of D1 and D2 mRNAs in distinct neuronal populations of the dorsal and ventral striatum. J Comp Neurol. 1995 May 8;355(3):418–426. doi: 10.1002/cne.903550308. [DOI] [PubMed] [Google Scholar]
  21. Li L., Yee C., Beavo J. A. CD3- and CD28-dependent induction of PDE7 required for T cell activation. Science. 1999 Feb 5;283(5403):848–851. doi: 10.1126/science.283.5403.848. [DOI] [PubMed] [Google Scholar]
  22. Liu H., Maurice D. H. Phosphorylation-mediated activation and translocation of the cyclic AMP-specific phosphodiesterase PDE4D3 by cyclic AMP-dependent protein kinase and mitogen-activated protein kinases. A potential mechanism allowing for the coordinated regulation of PDE4D activity and targeting. J Biol Chem. 1999 Apr 9;274(15):10557–10565. doi: 10.1074/jbc.274.15.10557. [DOI] [PubMed] [Google Scholar]
  23. Lobban M., Shakur Y., Beattie J., Houslay M. D. Identification of two splice variant forms of type-IVB cyclic AMP phosphodiesterase, DPD (rPDE-IVB1) and PDE-4 (rPDE-IVB2) in brain: selective localization in membrane and cytosolic compartments and differential expression in various brain regions. Biochem J. 1994 Dec 1;304(Pt 2):399–406. doi: 10.1042/bj3040399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. MacKenzie S. J., Baillie G. S., McPhee I., Bolger G. B., Houslay M. D. ERK2 mitogen-activated protein kinase binding, phosphorylation, and regulation of the PDE4D cAMP-specific phosphodiesterases. The involvement of COOH-terminal docking sites and NH2-terminal UCR regions. J Biol Chem. 2000 Jun 2;275(22):16609–16617. doi: 10.1074/jbc.275.22.16609. [DOI] [PubMed] [Google Scholar]
  25. MacKenzie S. J., Yarwood S. J., Peden A. H., Bolger G. B., Vernon R. G., Houslay M. D. Stimulation of p70S6 kinase via a growth hormone-controlled phosphatidylinositol 3-kinase pathway leads to the activation of a PDE4A cyclic AMP-specific phosphodiesterase in 3T3-F442A preadipocytes. Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3549–3554. doi: 10.1073/pnas.95.7.3549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morena A. R., Boitani C., de Grossi S., Stefanini M., Conti M. Stage and cell-specific expression of the adenosine 3',5' monophosphate-phosphodiesterase genes in the rat seminiferous epithelium. Endocrinology. 1995 Feb;136(2):687–695. doi: 10.1210/endo.136.2.7835302. [DOI] [PubMed] [Google Scholar]
  27. Oki N., Takahashi S. I., Hidaka H., Conti M. Short term feedback regulation of cAMP in FRTL-5 thyroid cells. Role of PDE4D3 phosphodiesterase activation. J Biol Chem. 2000 Apr 14;275(15):10831–10837. doi: 10.1074/jbc.275.15.10831. [DOI] [PubMed] [Google Scholar]
  28. Parvinen M. Regulation of the seminiferous epithelium. Endocr Rev. 1982 Fall;3(4):404–417. doi: 10.1210/edrv-3-4-404. [DOI] [PubMed] [Google Scholar]
  29. Polli J. W., Kincaid R. L. Expression of a calmodulin-dependent phosphodiesterase isoform (PDE1B1) correlates with brain regions having extensive dopaminergic innervation. J Neurosci. 1994 Mar;14(3 Pt 1):1251–1261. doi: 10.1523/JNEUROSCI.14-03-01251.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rajfer J., Aronson W. J., Bush P. A., Dorey F. J., Ignarro L. J. Nitric oxide as a mediator of relaxation of the corpus cavernosum in response to nonadrenergic, noncholinergic neurotransmission. N Engl J Med. 1992 Jan 9;326(2):90–94. doi: 10.1056/NEJM199201093260203. [DOI] [PubMed] [Google Scholar]
  31. Salanova M., Chun S. Y., Iona S., Puri C., Stefanini M., Conti M. Type 4 cyclic adenosine monophosphate-specific phosphodiesterases are expressed in discrete subcellular compartments during rat spermiogenesis. Endocrinology. 1999 May;140(5):2297–2306. doi: 10.1210/endo.140.5.6686. [DOI] [PubMed] [Google Scholar]
  32. Sasaki T., Kotera J., Yuasa K., Omori K. Identification of human PDE7B, a cAMP-specific phosphodiesterase. Biochem Biophys Res Commun. 2000 May 19;271(3):575–583. doi: 10.1006/bbrc.2000.2661. [DOI] [PubMed] [Google Scholar]
  33. Sassone-Corsi P. Transcription factors responsive to cAMP. Annu Rev Cell Dev Biol. 1995;11:355–377. doi: 10.1146/annurev.cb.11.110195.002035. [DOI] [PubMed] [Google Scholar]
  34. Sette C., Conti M. Phosphorylation and activation of a cAMP-specific phosphodiesterase by the cAMP-dependent protein kinase. Involvement of serine 54 in the enzyme activation. J Biol Chem. 1996 Jul 12;271(28):16526–16534. doi: 10.1074/jbc.271.28.16526. [DOI] [PubMed] [Google Scholar]
  35. Snyder G. L., Fienberg A. A., Huganir R. L., Greengard P. A dopamine/D1 receptor/protein kinase A/dopamine- and cAMP-regulated phosphoprotein (Mr 32 kDa)/protein phosphatase-1 pathway regulates dephosphorylation of the NMDA receptor. J Neurosci. 1998 Dec 15;18(24):10297–10303. doi: 10.1523/JNEUROSCI.18-24-10297.1998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Soderling S. H., Bayuga S. J., Beavo J. A. Cloning and characterization of a cAMP-specific cyclic nucleotide phosphodiesterase. Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8991–8996. doi: 10.1073/pnas.95.15.8991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Soderling S. H., Bayuga S. J., Beavo J. A. Isolation and characterization of a dual-substrate phosphodiesterase gene family: PDE10A. Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):7071–7076. doi: 10.1073/pnas.96.12.7071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Soderling S. H., Beavo J. A. Regulation of cAMP and cGMP signaling: new phosphodiesterases and new functions. Curr Opin Cell Biol. 2000 Apr;12(2):174–179. doi: 10.1016/s0955-0674(99)00073-3. [DOI] [PubMed] [Google Scholar]
  39. Surmeier D. J., Bargas J., Hemmings H. C., Jr, Nairn A. C., Greengard P. Modulation of calcium currents by a D1 dopaminergic protein kinase/phosphatase cascade in rat neostriatal neurons. Neuron. 1995 Feb;14(2):385–397. doi: 10.1016/0896-6273(95)90294-5. [DOI] [PubMed] [Google Scholar]
  40. Xu L. X., Tanaka Y., Bonderenko V. A., Matsuura I., Matsumoto H., Yamazaki A., Hayashi F. Phosphorylation of the gamma subunit of the retinal photoreceptor cGMP phosphodiesterase by the cAMP-dependent protein kinase and its effect on the gamma subunit interaction with other proteins. Biochemistry. 1998 Apr 28;37(17):6205–6213. doi: 10.1021/bi973087i. [DOI] [PubMed] [Google Scholar]
  41. Yarwood S. J., Steele M. R., Scotland G., Houslay M. D., Bolger G. B. The RACK1 signaling scaffold protein selectively interacts with the cAMP-specific phosphodiesterase PDE4D5 isoform. J Biol Chem. 1999 May 21;274(21):14909–14917. doi: 10.1074/jbc.274.21.14909. [DOI] [PubMed] [Google Scholar]
  42. Yuasa K., Kotera J., Fujishige K., Michibata H., Sasaki T., Omori K. Isolation and characterization of two novel phosphodiesterase PDE11A variants showing unique structure and tissue-specific expression. J Biol Chem. 2000 Oct 6;275(40):31469–31479. doi: 10.1074/jbc.M003041200. [DOI] [PubMed] [Google Scholar]
  43. Zhao A. Z., Zhao H., Teague J., Fujimoto W., Beavo J. A. Attenuation of insulin secretion by insulin-like growth factor 1 is mediated through activation of phosphodiesterase 3B. Proc Natl Acad Sci U S A. 1997 Apr 1;94(7):3223–3228. doi: 10.1073/pnas.94.7.3223. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES