Role of Yes kinase during early zebrafish development.
We have identified the Yes kinase in zebrafish eggs and investigated its role in development of the zebrafish embryo. In situ hybridization as well as immunofluorescence techniques demonstrated that Yes kinase is maternally expressed and is localized to the cortical region of the unfertilized egg. Fertilization resulted in concentration of Yes kinase to the blastodisc where it continued to be localized to the blastoderm cells through cleavage, gastrulation, and later development. Yes kinase activity was found to decrease abruptly at fertilization, then increase progressively during epiboly, and was maintained at high levels throughout gastrulation. The role of Yes kinase in development was tested by treating embryos with chemical protein tyrosine kinase (PTK) inhibitors such as 4-amino-5-(4-chlorophenyl)-7-(t-butyl) pyrazolo[3,4-d] pyrimidine (PP2) and by injection of antisense morpholinos. Both treatments resulted in the arrest of development at the beginning of the epiboly. Co-immunoprecipitation studies demonstrated that Yes kinase participates in a stable complex with focal adhesion kinase (FAK), which is phosphorylated in vitro. These results demonstrate that Yes kinase plays an important role in epiboly and indicate that Yes kinase participates in signaling by focal adhesion kinase during early development.
Tsai
Wen-Bin
WB
Department of Anatomy and Cell Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA.
Zhang
Xiaoming
X
Sharma
Dipika
D
Wu
Wenjun
W
Kinsey
William H
WH
D000595:Amino Acid Sequence; D000818:Animals; D004625:Embryo, Nonmammalian / Q000737:chemistry; D047108:Embryonic Development; D005260:Female; D008969:Molecular Sequence Data; D010063:Ovum / Q000737:chemistry; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000502:physiology*; D051575:Proto-Oncogene Proteins c-yes; D015027:Zebrafish / Q000196:embryology*; D029961:Zebrafish Proteins; D019061:src-Family Kinases / Q000032:analysis / Q000502:physiology*
D011518:Proto-Oncogene Proteins; D029961:Zebrafish Proteins; D051575:Proto-Oncogene Proteins c-yes; D019061:src-Family Kinases; C496640:yes1 protein, zebrafish