Hornbeck P, Huang K P, Paul W E
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1988 Apr;85(7):2279-83. doi: 10.1073/pnas.85.7.2279.
Lamin B was shown to be a major substrate of cellular phosphorylation in the response of lymphocytes to phorbol esters. Lamins A and C, which were not observed in lymphocytes, were also substrates of phorbol-stimulated phosphorylation in those cell types that express them. Lamin B phosphopeptides labeled with 32P in intact cells treated with phorbol 12-myristate 13-acetate were compared to those produced by in vitro phosphorylation with protein kinase M, cAMP-dependent protein kinase, and Ca2+/calmodulin-dependent protein kinase II. The phosphopeptides labeled by in vivo stimulation with phorbol esters are very similar to those phosphorylated in vitro by protein kinase M, a catalytic domain of protein kinase C. Phorbol treatment of interphase cells significantly reduces the amount of detergent-insoluble lamin B, suggesting that phosphorylation of lamin may alter the architecture of the nuclear lamina. In addition, we have shown that treatment of a B-cell line with antibodies to IgM induces a modest increase in lamin B phosphorylation. These results strongly suggest that ligands that are known to activate protein kinase C at the cell surface or in the cytosol also lead to the activation of a nuclear kinase activity with a protein kinase C-type specificity.
在淋巴细胞对佛波酯的反应中,核纤层蛋白B被证明是细胞磷酸化的主要底物。在淋巴细胞中未观察到的核纤层蛋白A和C,在表达它们的细胞类型中也是佛波酯刺激的磷酸化底物。将用佛波醇12-肉豆蔻酸酯13-乙酸酯处理的完整细胞中用32P标记的核纤层蛋白B磷酸肽与用蛋白激酶M、cAMP依赖性蛋白激酶和Ca2+/钙调蛋白依赖性蛋白激酶II进行体外磷酸化产生的磷酸肽进行比较。用佛波酯进行体内刺激标记的磷酸肽与蛋白激酶M(蛋白激酶C的催化结构域)在体外磷酸化的磷酸肽非常相似。用佛波醇处理间期细胞会显著减少去污剂不溶性核纤层蛋白B的量,这表明核纤层蛋白的磷酸化可能会改变核纤层的结构。此外,我们已经表明,用抗IgM抗体处理B细胞系会导致核纤层蛋白B磷酸化适度增加。这些结果强烈表明,已知在细胞表面或细胞质中激活蛋白激酶C的配体也会导致具有蛋白激酶C型特异性的核激酶活性的激活。