THE CDNA OF A HUMAN LYMPHOCYTE CYCLIC-AMP PHOSPHODIESTERASE (PDE IV) REVEALS A MULTIGENE FAMILY

Five protein families are needed to encompass the diversity of cyclic-AMP (cAMP) phosphodiesterases (PDE). Family IV PDEs (PDE IV) specifically hydrolyze cAMP with a low K(m), and are selectively inhibited by rolipram (Rp) and related drugs. Cloned cDNAs from rat (r) suggest that the PDE IV family comprises four distinct members, designated A, B, C and D. Using RNA from a human lymphocytic B-cell line (43D-Cl2), we have isolated a 3.8-kb cDNA by low-stringency screening using a rat PDE IV member B (r-PDE IV(B)) probe. Expression of the human (h) cDNA in Escherichia coli results in cAMP-specific PDE activity that is Rp sensitive. A single large open reading frame (ORF) predicts a 564-amino-acid protein with 92.9% identity to r-PDE IV(B); at the nucleotide level the identity is 86.3%. This h-PDE IV(B) clone, HPB106, differs from a related cDNA clone isolated by others from h-monocytes [Livi et al., Mol. Cell. Biol. 10 (1990) 2678-2686]. Our analysis identifies the monocyte clone with r-PDE IV(A). Southern blots using a 1.2-kb h-PDE IV(B) probe at low stringency suggest the presence of additional uncloned human PDE IV family members. Analysis of genomic Southern blots using short specific probes from the h-PDE IV(A) and h-PDE IV(B) cDNAs indicates that distinct genes encode these two PDE IV family members. RNA from fractionated normal human leukocytes shows major specific messages of 3.0 and 4.6 kb for h-PDE IV(A) and 3.7 kb for h-PDE IV(B). Comparison of our cDNA clones, and PCR analysis of cellular mRNA, suggests that alternative 5'-end sequences of h-PDE IV(B) are utilized. These results demonstrate a phenomenal complexity in the expression of human PDE IVs which suggests that unique PDE IV isozymes may control cAMP concentrations in cells.