Cyclic nucleotide phosphodiesterase of Dictyostelium discoideum and its glycoprotein inhibitor: Structure and expression of their genes
- 1 January 1991
- journal article
- research article
- Published by Wiley in Developmental Genetics
- Vol. 12 (1-2), 104-112
- https://doi.org/10.1002/dvg.1020120118
Abstract
The genes coding for the cyclic nucleotide phosphodiesterase (PD) and the PD inhibitory glycoprotein (PDI) have been cloned and characterized. The PDI gene was isolated as a 1.6 kb genomic fragment, which included the coding sequence containing two small introns and 510 nucleotides of non-translated 5′ sequence. From the deduced amino acid sequence we predict a protein with a molecular weight (MW) of 26,000 that, in agreement with previous data, contains 15% cysteine residues. Genomic Southern blot analysis indicates that only one gene encodes the inhibitor. Northern blot analysis shows a single transcript of 0.95 kb. The PDI gene is expressed early in development with little transcript remaining following aggregation. The appearance of PDI mRNA is prevented by the presence of cAMP, but when cAMP is removed the transcript appears within 30 minutes. When cAMP is applied to cells expressing PDI the transcript disappears with a half-life of less than 30 minutes. The PD gene of D. discoideum is transcribed into three mRNAs: a 1.9 kb mRNA specific for growth, a 2.4 kb mRNA specific for aggregation, and a 2.2 kb mRNA specific for late development. The 2.2 kb mRNA is also specific for prestalk cells, and is induced by differentiation-inducing factor. All three mRNAs contain the same coding sequence, and differ only in their 5′ non-coding sequences. Each mRNA is transcribed from a different promoter, and by using the chloramphenicol acyltransferase gene as a reporter, we have shown that each promoter displays the same regulation as its cognate mRNA. Transformation of wild-type strains with the PD gene causes PD overexpression which accelerates aggregation and blocks subsequent cell differentiation and pattern formation.Keywords
This publication has 62 references indexed in Scilit:
- Rescue of a Dictyostelium discoideum mutant defective in cyclic nucleotide phosphodiesteraseDevelopmental Biology, 1989
- The expression of two transcripts of the phosphodiesterase gene during the development of Dictyostelium discoideumDevelopmental Biology, 1987
- Physicochemical and hydrodynamic characterization of P-57, a neurospecific calmodulin binding proteinBiochemistry, 1986
- Characterization of the cyclic AMP phosphodiesterase activity secreted during development of Dictyostelium discoideumBiochimica et Biophysica Acta (BBA) - Protein Structure and Molecular Enzymology, 1983
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979
- Independent regulation of the extracellular cyclic AMP phosphodiesterase-inhibitor system and membrane differentiation by exogenous cyclic AMP in Dictyostelium discoideumDevelopmental Biology, 1978
- Cyclic AMP, Cyclic AMP Phosphodiesterase, and the Duration of the Interphase in Dictyostelium discoideumDifferentiation, 1977
- Defective cyclic adenosine-3′,5′-phosphate-phosphodiesterase regulation in morphogenetic mutants of Dictyostelium discoideumJournal of Molecular Biology, 1973
- Cyclic AMP phosphodiesterase interaction with its inhibitor of the slime mold, Dictyostelium discoideumBiochemical and Biophysical Research Communications, 1972
- Regulation of extracellular cyclic-AMP-phosphodiesterase activity during development of dictyostelium discoideumBiochemical and Biophysical Research Communications, 1971