{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,10]],"date-time":"2026-06-10T16:34:59Z","timestamp":1781109299612,"version":"3.54.1"},"reference-count":0,"publisher":"National Academy of Sciences","issue":"5","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[1995,2,28]]},"abstract":"<jats:p>c-Mil is the avian homologue of the mammalian serine\/threonine kinase c-Raf-1. c-Mil\/Raf is a mediator of signal transduction leading to gene expression via the c-Jun DNA-binding site, AP-1. Here we show that c-Mil immunopurified from MC29-virus-transformed quail fibroblasts phosphorylates c-Jun in vitro near its N terminus (Ser-63 and -73). Furthermore, the viral oncogene product Gag-Mil of the avian wild-type retrovirus MH2 phosphorylates c-Jun in vitro. A contribution by other known kinases phosphorylating c-Jun, such as the mitogen-activated protein kinases (MAPKs) and the c-Jun N-terminal kinases, was excluded by control reactions. c-Raf-1 and c-Jun directly interact in vitro as shown by various immobilized glutathione S-transferase-Raf fusion proteins which specify the cysteine-rich region of c-Mil\/Raf as the major N-terminal binding site. An additional minor binding site is located in the C-terminal region. The biological relevance of these results is demonstrated by coimmunoprecipitation of c-Jun and c-Mil from 32P-labeled MC29- and MH2-transformed fibroblasts as well as normal quail embryo fibroblasts, whereby c-Jun was identified by tryptic phosphopeptide analysis. The complexed c-Jun exhibits a decreased electrophoretic mobility corresponding to a more highly phosphorylated state. Cell fractionation analyses indicate that the c-Mil\/c-Jun complex is located in the cytoplasm. The data demonstrate that c-Jun can be a direct target of the protein kinase c-Mil\/Raf, suggesting an alternative pathway, which leads to c-Jun phosphorylation independent of the MAPKs and MAPK-related proteins.<\/jats:p>","DOI":"10.1073\/pnas.92.5.1421","type":"journal-article","created":{"date-parts":[[2006,5,31]],"date-time":"2006-05-31T09:27:59Z","timestamp":1149067679000},"page":"1421-1425","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":11,"title":["Direct interaction and N-terminal phosphorylation of c-Jun by c-Mil\/Raf."],"prefix":"10.1073","volume":"92","author":[{"given":"G","family":"Radziwill","sequence":"first","affiliation":[{"name":"Institut f\u00fcr Medizinische Virologie, Universit\u00e4t Z\u00fcrich, Switzerland."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"M","family":"Niehof","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Medizinische Virologie, Universit\u00e4t Z\u00fcrich, Switzerland."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C","family":"Rommel","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Medizinische Virologie, Universit\u00e4t Z\u00fcrich, Switzerland."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"K","family":"Moelling","sequence":"additional","affiliation":[{"name":"Institut f\u00fcr Medizinische Virologie, Universit\u00e4t Z\u00fcrich, Switzerland."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[1995,2,28]]},"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.92.5.1421","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,4,13]],"date-time":"2022-04-13T13:52:15Z","timestamp":1649857935000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/full\/10.1073\/pnas.92.5.1421"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1995,2,28]]},"references-count":0,"journal-issue":{"issue":"5","published-print":{"date-parts":[[1995,2,28]]}},"alternative-id":["10.1073\/pnas.92.5.1421"],"URL":"https:\/\/doi.org\/10.1073\/pnas.92.5.1421","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[1995,2,28]]},"assertion":[{"value":"1995-02-28","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}