Identification of V23RalA-Ser194 as a Critical Mediator for Aurora-A-induced Cellular Motility and Transformation by Small Pool Expression Screening

Autor: Chi Ying F. Huang, Si Jie Tsai, Chiun Jye Yuan, Jiunn Chyi Wu, Jung Mao Hsu, Chen-Kung Chou, Ming Jer Tang, Wey Jinq Lin, Tzong Yueh Chen, Chang Tze R. Yu
Rok vydání: 2005
Předmět:
Recombinant Fusion Proteins
DNA Mutational Analysis
Molecular Sequence Data
Motility
Cell Cycle Proteins
macromolecular substances
Protein Serine-Threonine Kinases
Xenopus Proteins
Biology
Kidney
Transfection
Biochemistry
Cell Line
Substrate Specificity
Mice
Dogs
Aurora Kinases
Cell Movement
Escherichia coli
Animals
Aurora Kinase B
Humans
Aurora Kinase C
Protein phosphorylation
Amino Acid Sequence
Cloning
Molecular
Phosphorylation
Molecular Biology
Aurora Kinase A
Sequence Homology
Amino Acid
Kinase
Cell migration
Cell Biology
Recombinant Proteins
RALA
Rats
Cell biology
enzymes and coenzymes (carbohydrates)
Protein kinase domain
embryonic structures
Mutagenesis
Site-Directed
ral GTP-Binding Proteins
Ectopic expression
biological phenomena
cell phenomena
and immunity
Signal transduction
Protein Kinases
Sequence Alignment
Zdroj: Journal of Biological Chemistry. 280:9013-9022
ISSN: 0021-9258
Popis: Human Aurora kinases have three gene family members: Aurora-A, Aurora-B, and Aurora-C. It is not yet established what the specificity of these kinases are and what signals relayed by their reactions. Therefore, we employed small pool expression screening to search for downstream substrates of Aurora-A. Interestingly, all of the identified Aurora-A substrates were resistant to serve as substrates for Aurora-B or Aurora-C, suggesting that these Aurora family members may have distinct substrate specificity for propagation of diverse signaling pathways, even though they share a conserved catalytic kinase domain. Of the candidate substrates, Aurora-A could increase the functional activity of RalA. Mutational analysis revealed that RalA-Ser194 was the phosphorylation site for Aurora-A. Ectopic expression of V23RalA-WT could enhance collagen I-induced cell migration and anchorage-independent growth in Madin-Darby canine kidney (MDCK) Aurora-A stable cell lines. In contrast, overexpression of V23RalA-S194A in MDCK Aurora-A-stable cell lines abolished the intrinsic migration and transformation abilities of Aurora-A. To our knowledge, this is the first systematic search for the downstream substrates of Aurora-A kinase. Moreover, these results support the notion that Aurora-A may act in concert with V23RalA through protein phosphorylation on Ser194 to promote collagen I-induced cell motility and anchorage-independent growth in MDCK epithelial cells.
Databáze: OpenAIRE
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