Proteomic and microarray analyses of the Dictyostelium Zak1-GSK-3 signaling pathway reveal a role in early development.
GskA, the Dictyostelium GSK-3 orthologue, is modified and activated by the dual-specificity tyrosine kinase Zak1, and the two kinases form part of a signaling pathway that responds to extracellular cyclic AMP. We identify potential cellular effectors for the two kinases by analyzing the corresponding null mutants. There are proteins and mRNAs that are altered in abundance in only one or the other of the two mutants, indicating that each kinase has some unique functions. However, proteomic and microarray analyses identified a number of proteins and genes, respectively, that are similarly misregulated in both mutant strains. The positive correlation between the array data and the proteomic data is consistent with the Zak1-GskA signaling pathway's functioning by directly or indirectly regulating gene expression. The discoidin 1 genes are positively regulated by the pathway, while the abundance of the H5 protein is negatively regulated. Two of the targets, H5 and discoidin 1, are well-characterized markers for early development, indicating that the Zak1-GskA pathway plays a role in development earlier than previously observed.
Strmecki
Lana
L
Biochemistry Department, Oxford University, South Parks Rd., Oxford OX1 3QU, United Kingdom.
Bloomfield
Gareth
G
Araki
Tsuyoshi
T
Dalton
Emma
E
Skelton
Jason
J
Schilde
Christina
C
Harwood
Adrian
A
Williams
Jeffrey G
JG
Ivens
Al
A
Pears
Catherine
C
D000818:Animals; D015153:Blotting, Western; D000242:Cyclic AMP / Q000378:metabolism; D004023:Dictyostelium / Q000201:enzymology* / Q000254:growth & development; D015180:Electrophoresis, Gel, Two-Dimensional; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000502:physiology; D018503:Genome, Protozoan; D038362:Glycogen Synthase Kinase 3 / Q000235:genetics / Q000378:metabolism*; D046228:Microarray Analysis; D020411:Oligonucleotide Array Sequence Analysis; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism*; D020543:Proteome / Q000032:analysis*; D015398:Signal Transduction / Q000502:physiology*
D020543:Proteome; D000242:Cyclic AMP; D011505:Protein-Tyrosine Kinases; D038362:Glycogen Synthase Kinase 3