The phosphorylated C-terminus of cAR1 plays a role in cell-type-specific gene expression and STATa tyrosine phosphorylation.
cAMP receptors mediate some signaling pathways via coupled heterotrimeric G proteins, while others are G-protein-independent. This latter class includes the activation of the transcription factors GBF and STATa. Within the cellular mounds formed by aggregation of Dictyostelium, micromolar levels of cAMP activate GBF function, thereby inducing the transcription of postaggregative genes and initiating multicellular differentiation. Activation of STATa, a regulator of culmination and ecmB expression, results from cAMP receptor-dependent tyrosine phosphorylation and nuclear localization, also in mound-stage cells. During mound development, the cAMP receptor cAR1 is in a low-affinity state and is phosphorylated on multiple serine residues in its C-terminus. This paper addresses possible roles of cAMP receptor phosphorylation in the cAMP-mediated stimulation of GBF activity, STATa tyrosine phosphorylation, and cell-type-specific gene expression. To accomplish this, we have expressed cAR1 mutants in a strain in which the endogenous cAMP receptors that mediate postaggregative gene expression in vivo are deleted. We then examined the ability of these cells to undergo morphogenesis and induce postaggregative and cell-type-specific gene expression and STATa tyrosine phosphorylation. Analysis of cAR1 mutants in which the C-terminal tail is deleted or the ligand-mediated phosphorylation sites are mutated suggests that the cAR1 C-terminus is not essential for GBF-mediated postaggregative gene expression or STATa tyrosine phosphorylation, but may play a role in regulating cell-type-specific gene expression and morphogenesis. A mutant receptor, in which the C-terminal tail is constitutively phosphorylated, exhibits constitutive activation of STATa tyrosine phosphorylation in pulsed cells in suspension and a significantly impaired ability to induce cell-type-specific gene expression. The constitutively phosphorylated receptor also exerts a partial dominant negative effect on multicellular development when expressed in wild-type cells. These findings suggest that the phosphorylated C-terminus of cAR1 may be involved in regulating aspects of receptor-mediated processes, is not essential for GBF function, and may play a role in mediating subsequent development.
Briscoe
C
C
Section of Cell and Developmental Biology, Center for Molecular Genetics, University of California at San Diego, 9500 Gilman Drive, La Jolla, California 92093-0634, USA.
Moniakis
J
J
Kim
J Y
JY
Brown
J M
JM
Hereld
D
D
Devreotes
P N
PN
Firtel
R A
RA
D000818:Animals; D000242:Cyclic AMP / Q000494:pharmacology; D004268:DNA-Binding Proteins / Q000378:metabolism; D004023:Dictyostelium / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D051790:G-Box Binding Factors; D005786:Gene Expression Regulation; D008958:Models, Molecular; D009024:Morphogenesis / Q000187:drug effects; D009154:Mutation; D010766:Phosphorylation; D015800:Protozoan Proteins / Q000378:metabolism*; D011953:Receptors, Cyclic AMP / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D050791:STAT Transcription Factors; D017384:Sequence Deletion; D014157:Transcription Factors / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism
D004268:DNA-Binding Proteins; D051790:G-Box Binding Factors; D015800:Protozoan Proteins; D011953:Receptors, Cyclic AMP; D050791:STAT Transcription Factors; C426583:STATa protein, Dictyostelium; D014157:Transcription Factors; C068125:cyclic AMP receptor cAR1; D014443:Tyrosine; D000242:Cyclic AMP