The entire coding sequence of wild-type mouse p53 was expressed in Escherichia coli under control of the PL promoter of bacteriophage lambda. The bacterial p53 protein had identical mobility to p53 from SV3T3 cells on SDS polyacrylamide gels and was recognized in bacterial lysates by three p53-specific monoclonal antibodies, including PAb246 which is specific for wild-type mouse p53. Immunoprecipitates of the bacterial p53 were phosphorylated by a highly purified preparation of rat casein kinase II; the stoichiometry of incorporation was approximately 1 mol of phosphate per mol of p53. The phosphorylated residue was identified by phosphopeptide mapping as serine 389, which is a major site of p53 phosphorylation in vivo. p53 (serine 389) kinase activity was detected on lysates of SV3T3 cells; this activity co-purified with casein kinase II on phosphocellulose and Mono Q columns and was inhibited by heparin. Immunoprecipitates of the p53-T antigen complex from SV3T3 cells also had associated serine 389 kinase activity. Phosphorylation of serine 389 by this kinase was potently inhibited by heparin and quenched by excess unlabelled GTP. The data indicate that p53 is a physiological substrate of casein kinase II, which is stimulated in response to mitogens, phosphorylates nuclear oncoproteins, and may play a role in the transduction of extracellular signals to the nucleus.
|Number of pages||8|
|Publication status||Published - 1990|
Meek, D. W., Simon, S., Kikkawa, U., & Eckhart, W. (1990). The p53 tumour suppressor protein is phosphorylated at serine 389 by casein kinase II. EMBO Journal, 9(10), 3253-3260. http://www.scopus.com/results/results.url?sort=plf-f&src=s&st1=The+p53+tumour+suppressor+protein+is+phosphorylated+at+serine+389+by+casein+kinase+II&sid=BE13A0E576A9FA1B1123BA5975278CCE.zQKnzAySRvJOZYcdfIziQ%3a470&sot=b&sdt=b&sl=100&s=TITLE-ABS-KEY%28The+p53+tumour+suppressor+protein+is+phosphorylated+at+serine+389+by+casein+kinase+II%29&origin=searchbasic&txGid=BE13A0E576A9FA1B1123BA5975278CCE.zQKnzAySRvJOZYcdfIziQ%3a47