Cell-permeable non-hydrolyzable cAMP derivatives as tools for analysis of signaling pathways controlling gene regulation in Dictyostelium

A novel class of cAMP derivatives were tested for binding to surface cAMP receptors (CAR), protein kinase A (PKA), and cAMP-phosphodiesterase (PDE) and for induction of three classes of cAMP regulated genes in Dictyostelium discoideum. These derivatives carry sulfur substitutions for either the axia...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of biological chemistry 1993-03, Vol.268 (9), p.6323-6331
Main Authors: Schaap, P, Ments-Cohen, M. van, Soede, R.D.M, Brandt, R, Firtel, R.A, Dostmann, W, Genieser, H.G, Jastorff, B, Haastert, P.J.M. van
Format: Article
Language:English
Subjects:
3',5'-Cyclic-AMP Phosphodiesterases - metabolism
6-thioethyl-purineriboside 3',5'-monophosphorothioate
activation
ACTIVIDAD ENZIMATICA
ACTIVITE ENZYMATIQUE
ADENOSINE MONOPHOSPHATE
ADENOSINMONOFOSFATO
analogs
Animals
Biological and medical sciences
Cell Membrane Permeability
cell membranes
Cell physiology
CHIMIORECEPTEUR
cyclic AMP
Cyclic AMP - analogs & derivatives
Cyclic AMP - metabolism
Dictyostelium - genetics
Dictyostelium - metabolism
Dictyostelium discoideum
ESTERASAS
ESTERASE
EXPRESION GENICA
EXPRESSION DES GENES
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation
gene regulation
GENETICA
GENETIQUE
Kinetics
MIXOMICETES
Molecular and cellular biology
Molecular Structure
MYXOMYCETES
pathways
permeability
protein kinase A
Protein Kinases - metabolism
PROTEINA QUINASA
PROTEINE KINASE
QUIMIORECEPTORS
receptors
Receptors, Cyclic AMP - metabolism
Signal Transduction
Thionucleotides - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A novel class of cAMP derivatives were tested for binding to surface cAMP receptors (CAR), protein kinase A (PKA), and cAMP-phosphodiesterase (PDE) and for induction of three classes of cAMP regulated genes in Dictyostelium discoideum. These derivatives carry sulfur substitutions for either the axial (Sp) or equatorial (Rp) exocyclic oxygen atoms, while further modifications were introduced to provide specificity for binding to either CAR or PKA, and/or to increase lipophilicity and render the derivatives membrane-permeable. All derivatives bind weakly to PDE and are almost not degraded during incubation with Dictyostelium cells. One cAMP derivative, 6-thioethylpurineriboside 3',5'-monophosphorothioate, Sp-isomer (Sp-6SEtcPuMPS), fulfills the criteria for selective activation of PKA in vivo. The compound enters Dictyostelium cells and reaches an intracellular concentration of 1 micromolar, sufficient to activate PKA, at an extracellular concentration of 30 micromolar, which is insufficient to activate CAR. Expression of cAMP-regulated prespore and prestalk genes and the aggregative PDE gene are effectively induced by CAR agonists and very poorly by PKA agonists. Even Sp-6SEtcPuMPS is ineffective to induce gene expression. These data not only indicate that surface cAMP receptors are the first targets for cAMP-induced gene expression, but argue against direct induction of expression of these genes by cAMP-induced PKA activation
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)53256-7