. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. FERTILIZATION AND DEVELOPMENTAL BIOLOGY Reference: liinl. Bull. 191: 283-284. (October, 1996) 283 A DNA-Inducible Kinase Activity Undergoes a Change in Cellular Localization During Development in Arbacia Jvolshnabala Kanungo, James A. Walker*, David Woodrum, John A. Ilardin. and Howard Rasmussen (Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, GA 30912) DNA-dependent phosphorylation has been detected in vari- ous cell types from a wide range of organisms. The enzyme re- sponsible for this a
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. FERTILIZATION AND DEVELOPMENTAL BIOLOGY Reference: liinl. Bull. 191: 283-284. (October, 1996) 283 A DNA-Inducible Kinase Activity Undergoes a Change in Cellular Localization During Development in Arbacia Jvolshnabala Kanungo, James A. Walker*, David Woodrum, John A. Ilardin. and Howard Rasmussen (Institute of Molecular Medicine and Genetics, Medical College of Georgia, Augusta, GA 30912) DNA-dependent phosphorylation has been detected in vari- ous cell types from a wide range of organisms. The enzyme re- sponsible for this activity in human cell lines, DNA-dependent protein kinase (DNA-PK), has been well characterized. Hu- man DNA-PK is composed of two subunits—a regulatory sub- unit (Ku) and a catalytic subunit (p460, previously known as p350) and has been shown to have roles in transcription, re- combination and repair of DNA (1-4). A DNA-dependent kinase activity has previously been found in crude extracts from fertilized sea urchin eggs (5, 6). In vitro kinase assays have shown that this activity is present in cyto- plasmic extracts of fertilized Arbacia eggs only up to the 4-cell stage (6). In this report we further examine the cellular localiza- tion of DNA-dependent kinase activity in urchin development. We performed in vitro DNA-dependent kinase assays using a specific peptide derived from p53—a known substrate of hu- Department of Biochemistry, University of Cambridge. UK. man DNA-PK (7). This enabled us to compare the relative lev- els of activity in both cytoplasmic and whole cell extracts from different stages of development. As in the assays performed with a-casein as a substrate (6), we were unable to induce the kinase activity in extracts from unfertilized eggs, although a high level of DNA-inducible activity is present in both cyto- plasmic and whole cell extracts of fertilized eggs (Fig. la). This activity gradually decreases in cytoplasmic extracts of 2- and 4- cell stages (u
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