Linked Life Data

1326532

Source:http://linkedlifedata.com/resource/pubmed/id/1326532

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
26
pubmed:dateCreated
1992-10-19
pubmed:databankReference
pubmed:abstractText
Multiple isozymes of cyclic nucleotide phosphodiesterases (PDEs) are expressed simultaneously in mammalian tissues. To identify and clone these PDEs, a polymerase chain reaction (PCR) strategy was developed using degenerate oligonucleotide primers designed to hybridize with highly conserved PDE DNA domains. Both known and novel PDEs were cloned from rat liver, the mouse K30a-3.3 lymphoma cell line, and a human hypothalamus cDNA library, demonstrating that these PCR primers can be used to amplify the cDNA of multiple PDE isozymes. One unique mouse PDE clone was found to encode a polypeptide identical with the corresponding portion of the bovine brain 63-kDa calmodulin-dependent PDE as reported in the companion article (Bentley, J. K., Kadlecek, A., Sherbert, C. H., Seger, D., Sonnenburg, W. K., Charbonneau, H., Novack, J. P., and Beavo, J. A. (1992) J. Biol. Chem. 267, 18676-18682). This mouse clone was used as a probe to screen a rat brain cDNA library for a full-length clone. The conceptual translation of the nucleotide sequence of the resulting rat clone has an open reading frame of 535 amino acids and maintains a high degree of homology with the bovine 63-kDa calmodulin-dependent PDE, indicating that this protein is likely to be the rat homolog of the 63-kDa calmodulin-dependent PDE. Expression of the full-length clone in Escherichia coli yielded a cGMP hydrolyzing activity that was stimulated severalfold by calmodulin. Northern blot analysis demonstrated that the mRNA encoding this PDE is highly expressed in rat brain and also in the S49.1 T-lymphocyte cell line. These data demonstrate that the PCR method described is a viable strategy to isolate cDNA clones of known and novel members of different families of PDE isozymes. Molecular cloning of these PDEs will provide valuable tools for investigating the roles of these isozymes in regulation of intracellular concentrations of the cyclic nucleotides.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
267
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
18683-8
pubmed:dateRevised
2007-11-15
pubmed:meshHeading
pubmed-meshheading:1326532-3',5'-Cyclic-AMP Phosphodiesterases, pubmed-meshheading:1326532-3',5'-Cyclic-GMP Phosphodiesterases, pubmed-meshheading:1326532-Amino Acid Sequence, pubmed-meshheading:1326532-Animals, pubmed-meshheading:1326532-Base Sequence, pubmed-meshheading:1326532-Blotting, Northern, pubmed-meshheading:1326532-Cloning, Molecular, pubmed-meshheading:1326532-Cyclic Nucleotide Phosphodiesterases, Type 1, pubmed-meshheading:1326532-DNA, pubmed-meshheading:1326532-Isoenzymes, pubmed-meshheading:1326532-Liver, pubmed-meshheading:1326532-Mice, pubmed-meshheading:1326532-Molecular Sequence Data, pubmed-meshheading:1326532-Polymerase Chain Reaction, pubmed-meshheading:1326532-Rats, pubmed-meshheading:1326532-Rats, Inbred Strains, pubmed-meshheading:1326532-Sequence Homology, Nucleic Acid, pubmed-meshheading:1326532-Transcription, Genetic, pubmed-meshheading:1326532-Tumor Cells, Cultured
pubmed:year
1992
pubmed:articleTitle
A polymerase chain reaction strategy to identify and clone cyclic nucleotide phosphodiesterase cDNAs. Molecular cloning of the cDNA encoding the 63-kDa calmodulin-dependent phosphodiesterase.
pubmed:affiliation
Division of Pediatric Endocrinology, University of North Carolina, Chapel Hill 27599.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.