MOLECULAR-CLONING OF A NOVEL SPLICE VARIANT OF HUMAN TYPE-IVA (PDE-IVA) CYCLIC-AMP PHOSPHODIESTERASE AND LOCALIZATION OF THE GENE TO THE P13.1-Q12 REGION OF HUMAN-CHROMOSOME-19

We have isolated from a human T-cell Jurkat cDNA. library a novel human cDNA (2EL) that is closely related to the human type-IV PDE splice variant family 'A' (PDE-IVA) cDNA characterized previously by us [Sullivan, Egerton, Shakur, Marquardsen and Houslay (1994) Cell. Signalling 6, 793-812]; (h6.1, PDE-IVA/h6.1; HSPDE4A7). (PDE stands for cyclic nucleotide phosphodiesterase). The novel cDNA 2EL (PDE IVA/2EL; HSPDE4A8) contains two regions of unique sequence not found in PDE-IVA/h6.1. These are a distinct 5'-end and a 34 bp insert which occurs within a domain thought to encode the type-IV PDE catalytic site and which can be expected to result in premature truncation of any expressed protein. HSPDE4A8 appeared to be catalytically inactive. Isolation and characterization of a human genomic cosmid clone revealed that 2EL and h6.1 represent alternative splice variants of the human PDE-IVA gene. Using a unique sequence found at the 5'-end of the 2EL cDNA, a probe was generated which was used to screen the DNA of human-hamster hybrids. This located the human gene for PDE-IVA to human chromosome 19. Through both the analysis of genomic DNAs from a human-hamster somatic cell hybrid panel and also using fluorescent in situ hybridization, it was shown that the human PDE-IVA gene is located on human chromosome 19, between p13.1 and q12. This region on chromosome 19 has been shown to be related to genetic diseases such as the autosomal dominant cerebrovascular disease CADASIL, susceptibility to late-onset Alzheimer's disease and changes seen in benign pituitary and thyroid adenomas.