Challenge of human Jurkat T-cells with the adenylate cyclase activator forskolin elicits major changes in cAMP phosphodiesterase (PDE) expression by up-regulating PDE3 and inducing PDE4D1 and PDE4D2 splice variants as well as down-regulating a novel PDE4A splice variant

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dc.authoridHouslay, Miles/0000-0002-3826-8091
dc.contributor.authorErdogan, S
dc.contributor.authorHouslay, MD
dc.date.accessioned2024-09-18T21:05:05Z
dc.date.available2024-09-18T21:05:05Z
dc.date.issued1997
dc.departmentHatay Mustafa Kemal Üniversitesien_US
dc.description.abstractThe cAMP phosphodiesterase (PDE) 3 and PDE4 isoforms provide the major cAMP-hydrolysing PDE activities in Jurkat T-cells, with additional contributions from the PDE1 and PDE2 isoforms. Challenge of cells with the adenylate cyclase activator forskolin led to a rapid: albeit transient, increase in PDE3 activity occurring over the first 45 min, followed by a sustained increase in PDE3 activity which began after similar to 3 h and continued for at least 24 h. Only this second phase of increase in PDE3 activity was blocked by the transcriptional inhibitor actinomycin D. After similar to 3 h of exposure to forskolin, PDE4 activity had increased, via a process that could be inhibited by actinomycin D, and it remained elevated for at least a 24 h period. Such actions of forskolin were mimicked by cholera toxin and 8-bromo-cAMP. Forskolin increased intracellular cAMP concentrations in a time-dependent fashion and its action was enhanced when PDE induction was blocked with actinomycin D. Reverse transcription (RT)-PCR analysis, using generic primers designed to detect transcripts representing enzymically active products of the four PDE4 genes, identified transcripts for PDE4A and PDE4D but not for PDE4B or PDE4C in untreated Jurkat T-cells. Forskolin treatment did not induce transcripts for either PDE4B or PDE4C; however, it reduced the RT-PCR signal for PDE4A transcripts and markedly enhanced that for PDE4D transcripts. Using RT-PCR primers for PDE4 splice variants, a weak signal for PDE4D1 was evident in control cells whereas, in forskolin-treated cells, clear signals for both PDE4D1 and PDE4D2 were detected. RT-PCR analysis of the PDE4A species indicated that it was not the PDE4A isoform PDE-46 (PDE4A4B). Immunoblotting of control cells for PDE4 forms identified a single PDE4A species of similar to 118 kDa, which migrated distinctly from the PDE4A4B isoform PDE-46, with immunoprecipitation analyses showing that it provided all of the PDE4 activity in control cells. Forskolin treatment led to a marked decrease of this novel PDE4A species and allowed the detection of a strong signal for an similar to 67 kDa PDE4D species, suggested to be PDE4D1, but did not induce PDE4B and PDE4C isoforms. Elevation of intracellular cAMP concentrations in Jurkat T-cells thus exerts a highly selective effect on the transcriptional activity of the genes encoding the various PDE4 isoforms. This leads to the down-regulation of a novel PDE4A splice variant and the induction of PDE4D1 and PDE4D2 splice variants, leading to a net increase in the total PDE4 activity of Jurkat T-cells.en_US
dc.identifier.doi10.1042/bj3210165
dc.identifier.endpage175en_US
dc.identifier.issn0264-6021
dc.identifier.pmid9003416en_US
dc.identifier.scopus2-s2.0-0031028090en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.startpage165en_US
dc.identifier.urihttps://doi.org/10.1042/bj3210165
dc.identifier.urihttps://hdl.handle.net/20.500.12483/13346
dc.identifier.volume321en_US
dc.identifier.wosWOS:A1997WB61900021en_US
dc.identifier.wosqualityQ1en_US
dc.indekslendigikaynakWeb of Scienceen_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherPortland Pressen_US
dc.relation.ispartofBiochemical Journalen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectCyclic-Amp Phosphodiesteraseen_US
dc.subjectDependent Protein-Kinaseen_US
dc.subjectLiver Plasma-Membranesen_US
dc.subjectNucleotide Phosphodiesterasesen_US
dc.subjectMolecular-Cloningen_US
dc.subjectHormonal-Regulationen_US
dc.subjectMessenger-Rnaen_US
dc.subjectInhibitionen_US
dc.subjectGeneen_US
dc.subjectLocalizationen_US
dc.titleChallenge of human Jurkat T-cells with the adenylate cyclase activator forskolin elicits major changes in cAMP phosphodiesterase (PDE) expression by up-regulating PDE3 and inducing PDE4D1 and PDE4D2 splice variants as well as down-regulating a novel PDE4A splice varianten_US
dc.typeArticleen_US

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