Identification of anisomycin-activated kinases p45 and p55 in murine cells as MAPKAP kinase-2

Eva Cano, Yair N. Doza, Rachel Ben-Levy, Philip Cohen, Louis C. Mahadevan

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    Abstract

    We recently showed that EGF and anisomycin activate two kinases, p45 and p55, whose distinguishing feature is that their detection in in-gel kinase assays is enhanced by copolymerised poly-Glu/Tyr or poly-Glu/Phe. Their activation characteristics and sizes are strikingly similar to those of JNK/SAPKs, which are also strongly activated by anisomycin. However, we show here that p45 and p55 are not JNK/SAPKs but murine forms of MAPKAP kinase-2 because: (i) Detection of immunoprecipitated JNK/SAPKs is completely dependent on the presence of c-Jun as substrate in the in-gel kinase assays, whereas detection of p45 and p55 is not. (ii) Detection of p45 and p55 activity is enhanced by the presence of poly-Glu/Tyr or poly-Glu/Phe, whereas JNK/SAPKs are not detectable under these conditions. (iii) Although the sizes of the murine JNK/SAPKs and MAPKAP K-2 are similar, human JNK/SAPKs migrate at 45 and 55 kDa whereas human MAPKAP K-2 migrates at 50 kDa; the poly-Glu/Tyr-enhanced activity in human cells migrates at 50 kDa. (iv) Purified rabbit muscle MAPKAP K-2 is detectable as two bands of activity on in-gel kinase assays and their detection is enhanced by poly-Glu/Tyr. (v) Finally, the anisomycin-activated poly-Glu/Tyr-enhanced p45 and p55 kinases can be immunoprecipitated from murine cells using an anti-MAPKAP K-2 antibody. Thus, EGF- and anisomycin-activated p45 and p55 are not JNK/SAPKs but MAPKAP K-2, implying that both these agents activate the p38/RK MAP kinase cascade.

    Original languageEnglish
    Pages (from-to)805-812
    Number of pages8
    JournalOncogene
    Volume12
    Issue number4
    Publication statusPublished - 13 Mar 1996

    Keywords

    • Anisomycin
    • Growth-factors
    • Intracellular signalling
    • MAP kinases
    • MAPKAP kinase-2
    • Stress kinases

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    Cano, E., Doza, Y. N., Ben-Levy, R., Cohen, P., & Mahadevan, L. C. (1996). Identification of anisomycin-activated kinases p45 and p55 in murine cells as MAPKAP kinase-2. Oncogene, 12(4), 805-812.