Isolation and characterization of a previously undetected human cAMP phosphodiesterase by complementation of cAMP phosphodiesterase- deficient Saccharomyces cerevisiae

Michaeli, T., Bloom, T. J., Martins, T., Loughney, K., Ferguson, K., Riggs, M., Rodgers, L., Beavo, J. A., Wigler, M. H. (June 1993) Isolation and characterization of a previously undetected human cAMP phosphodiesterase by complementation of cAMP phosphodiesterase- deficient Saccharomyces cerevisiae. Journal of Biological Chemistry, 268 (17). pp. 12925-12932. ISSN 0021-9258

URL: https://www.ncbi.nlm.nih.gov/pubmed/8389765

Abstract

We have established a highly sensitive functional screen for the isolation of cDNAs encoding cAMP phosphodiesterases (PDEs) by complementation of defects in a Saccharomyces cerevisiae strain lacking both endogenous cAMP PDE genes, PDE1 and PDE2. Three groups of cDNAs corresponding to three distinct human genes encoding cAMP-specific PDEs were isolated from a human glioblastoma cDNA library using this functional screen. Two of these genes are closely related to the Drosophila dunce cAMP-specific PDE. The third gene, which we named HCP1, encoded a novel cAMP-specific PDE. HCP1 has an amino acid sequence related to the sequences of the catalytic domains of all cyclic nucleotide PDEs. HCP1 is a high affinity cAMP-specific PDE (K(m) = 0.2 muM) that does not share other properties of the cAMP-specific PDE family, i.e. extensive sequence homology to the Drosophila dunce cAMP PDE and sensitivity to rolipram and R020-1724. The PDE activity of HCP1 is not sensitive to cGMP or other inhibitors of the cGMP-inhibitable PDEs, such as milrinone. The biochemical and pharmacological properties of HCP1 suggest that it is a member of a previously undiscovered cyclic nucleotide PDE family. Northern blot analysis indicates that high levels of HCP1 mRNA are present in human skeletal muscle.

Item Type: Paper
Uncontrolled Keywords: CYCLIC-NUCLEOTIDE PHOSPHODIESTERASE cyclic nucleotide phosphodiesterase AMP PHOSPHODIESTERASE phosphodiesterase MOLECULAR-CLONING molecular cloning BAKERS-YEAST Bakers yeast GENE gene PURIFICATION purification EXPRESSION expression INHIBITORS inhibitors ACTIVATION activation HOMOLOGS homologs
Subjects: organism description > yeast > Saccharomyces
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > cDNA
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function > gene expression
CSHL Authors:
Communities: CSHL labs > Wigler lab
Depositing User: CSHL Librarian
Date: June 1993
Date Deposited: 12 Apr 2012 13:37
Last Modified: 07 Nov 2016 20:39
Related URLs:
URI: http://repository.cshl.edu/id/eprint/26230

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