Characterization of five different proteins produced by alternatively spliced mRNAs from the human cAMP-specific phosphodiesterase PDE4D gene

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dc.authorscopusid16188193800
dc.authorscopusid7004540505
dc.authorscopusid57197987128
dc.authorscopusid56627340100
dc.authorscopusid6603407836
dc.authorscopusid57213207538
dc.authorscopusid58397674100
dc.contributor.authorBolger, Graeme B.
dc.contributor.authorErdogan, Suat
dc.contributor.authorJones, Randy E.
dc.contributor.authorLoughney, Kate
dc.contributor.authorScotland, Grant
dc.contributor.authorHoffmann, Ralf
dc.contributor.authorWilkinson, Ian
dc.date.accessioned2024-09-19T15:48:46Z
dc.date.available2024-09-19T15:48:46Z
dc.date.issued1997
dc.departmentHatay Mustafa Kemal Üniversitesien_US
dc.description.abstractWe have isolated and characterized complete cDNAs for two isoforms (HSPDE4D4 and HSPDE4A5) encoded by the human PDE4D gene, one of four genes that encode cAMP-specific rolipram-inhibited 3',5'-cyclic nucleotide phosphodiesterases (type IV PDEs; PDE4 family). The HSPDE4D4 and HSPDE4D5 cDNAs encode proteins of 810 and 746 amino acids respectively. A comparison of the nucleotide sequences of these two cDNAs with those encoding the three other human PDE4D proteins (HSPDE4D1, HSPDE4D2 and HSPDE4D3) demonstrates that each corresponding mRNA transcript has a unique region of sequence at or near its 5'-end, consistent with alternative mRNA splicing. Transient expression of the five cDNAs in monkey COS-7 cells produced proteins of apparent molecular mass under denaturing conditions of 68, 68, 95, 119 and 105 kDa for isoforms HSPDE4D1-5 respectively. Immunoblotting of human cell lines and rat brain demonstrated the presence of species that co-migrated with the proteins produced in COS-7 cells. COS-cell-expressed and native HSPDE4D1 and HSPDE4D2 were found to exist only in the cytosol, whereas HSPDE4D3, HSPDE4D4 and HSPDE4D5 were found in both cytosolic and particulate fractions. The IC50 values for the selective PDE4 inhibitor rolipram for the cytosolic forms of the five enzymes were similar (0.05-0.14 ?M), whereas they were 2-7-fold higher for the particulate forms of HSPDE4D3 and HSPDE4D5 (0.32 and 0.59 ?M respectively), than for the corresponding cytosolic forms. Our data indicate that the N-terminal regions of the HSPDE4D3, HSPDE4D4 and HSPDE4D5 proteins, which are derived from alternatively spliced regions of their mRNAs, are important in determining their subcellular localization, activity and differential sensitivity to inhibitors.en_US
dc.description.sponsorshipNational Cancer Institute, NCI, (P30CA042014)en_US
dc.identifier.doi10.1042/bj3280539
dc.identifier.endpage548en_US
dc.identifier.issn0264-6021
dc.identifier.issue2en_US
dc.identifier.pmid9371713en_US
dc.identifier.scopus2-s2.0-0030665303en_US
dc.identifier.scopusqualityQ1en_US
dc.identifier.startpage539en_US
dc.identifier.urihttps://doi.org/10.1042/bj3280539
dc.identifier.urihttps://hdl.handle.net/20.500.12483/15297
dc.identifier.volume328en_US
dc.indekslendigikaynakScopusen_US
dc.indekslendigikaynakPubMeden_US
dc.language.isoenen_US
dc.publisherPortland Press Ltden_US
dc.relation.ispartofBiochemical Journalen_US
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanıen_US
dc.rightsinfo:eu-repo/semantics/openAccessen_US
dc.subjectAnimaliaen_US
dc.subjectcomplementary DNAen_US
dc.subjectcyclic AMPen_US
dc.subjectmessenger RNAen_US
dc.subjectphosphodiesteraseen_US
dc.subjectproteinen_US
dc.subjectrolipramen_US
dc.subjectalternative RNA splicingen_US
dc.subjectamino terminal sequenceen_US
dc.subjectanimal cellen_US
dc.subjectarticleen_US
dc.subjectbrainen_US
dc.subjectcomparative studyen_US
dc.subjectcytosolen_US
dc.subjectDNA determinationen_US
dc.subjectgene expressionen_US
dc.subjecthumanen_US
dc.subjecthuman cellen_US
dc.subjectimmunoblottingen_US
dc.subjectmolecular weighten_US
dc.subjectnonhumanen_US
dc.subjectnucleotide sequenceen_US
dc.subjectpriority journalen_US
dc.subjectprotein analysisen_US
dc.subjectprotein denaturationen_US
dc.subjectprotein expressionen_US
dc.subjectprotein localizationen_US
dc.subjectraten_US
dc.titleCharacterization of five different proteins produced by alternatively spliced mRNAs from the human cAMP-specific phosphodiesterase PDE4D geneen_US
dc.typeArticleen_US

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