. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 282 REPORTS FROM THE MBL GENERAL SCIENTIFIC MEETINGS + + DNA DNA. c Pluteus tic i SO' Post-Pert. E UPC 103' Post-Pert. Figure 1. Autontdiogiam v//mrv variation in DNA-dependent protein phosphorylation activity during embryonic development. Cyloplasmic extract-, from eggi and einhiyoi were prepared already reported(4). A 15 »l reaction required 10 pi ofcyloplasmic extract, 75 ng of sonicated calf t hymns DNA, 15 ng dephosphorylated ct-casein (Sigma) as substrate, 2 in At A/#('/_*, and 130 pM ATP. The reaction was carried
. The Biological bulletin. Biology; Zoology; Biology; Marine Biology. 282 REPORTS FROM THE MBL GENERAL SCIENTIFIC MEETINGS + + DNA DNA. c Pluteus tic i SO' Post-Pert. E UPC 103' Post-Pert. Figure 1. Autontdiogiam v//mrv variation in DNA-dependent protein phosphorylation activity during embryonic development. Cyloplasmic extract-, from eggi and einhiyoi were prepared already reported(4). A 15 »l reaction required 10 pi ofcyloplasmic extract, 75 ng of sonicated calf t hymns DNA, 15 ng dephosphorylated ct-casein (Sigma) as substrate, 2 in At A/#('/_*, and 130 pM ATP. The reaction was carried out at 15°C. and started hy adding 10 »Ci of \gumma-''/']!//' (MHH) Ci/mmol) (NEN, Du Pont). The phosphorylation reaction (I nD was analyzed on a 10% polyacrylamide gel (SDS-PAGE) i,—e \lracl only (without any exogenous .<iiih.\iruie); C—extract with exogenous a-casein added as an exogenous . rate. Arrows indicate the position ofphosphorylated casein. Absence and presence ofdsDNA in the extract are indicated hy— and + , respectively Therefore, a specific substrate is needed to assay for the specific enzyme activity in whole cell extracts. These results demonstrate the presence of a DNA-dependent protein kinase in the cytoplasm of eggs and embryos of sea ur- chin. The rapid appearance of activity in the cytoplasmic ex- tracts from fertilized eggs could be due to either the synthesis of the enzyme itself or the activation of preexisting inactive en- zyme by post-translational modification. However, we can not rule out the possibility that the enzyme is nuclear in unfertil- ized eggs and is released after fertilization. We thank Dr. Carl Anderson and Dr. William Dvnan for helpful discussions. Grant support came from NIH (AR 32549 to JAH), Mason Trust, the Arthritis Foundation. Literature Cited 1. Carter, I., I. Vancurova, I. Sun, \V. Lou, and S. DeLeon. 1990. Mol Cell Biol. 10:6460-6471. 2 Finnic, N. J., T. M. Gottlieb, T. Blunt, P. A. Jeggo, and S. P. Jack- son
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