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The ability to alter the cellular signaling pathways suggests that Nef can associate with signaling proteins. In the present report, we show that Nef can interact with calmodulin, the major intracellular receptor for calcium. Coimmunoprecipitation analyses with lysates from the NIH3T3 cell line constitutively expressing the native HIV\u20101 Nef protein revealed the presence of a stable Nef\u2010calmodulin complex. When lysates from NIH3T3 cells were incubated with calmodulin\u2010agarose beads in the presence of CaCl<jats:sub>2<\/jats:sub> or EGTA, calcium ion drastically enhanced the interaction between Nef and calmodulin, suggesting that the binding is under the influence of Ca<jats:sup>2+<\/jats:sup> signaling. Glutathione <jats:italic>S<\/jats:italic>\u2010transferase\u2010Nef fusion protein bound directly to calmodulin with high affinity. Using synthetic peptides based on the N\u2010terminal sequence of Nef, we determined that within a 20\u2010amino\u2010acid N\u2010terminal basic domain was sufficient for calmodulin binding. Furthermore, the myristoylated peptide bound to calmodulin with higher affinity than nonmyris\u2010toylated form. Thus, the N\u2010terminal myristoylation domain of Nef plays an important role in interacting with calmodulin. This domain is highly conserved in several HIV\u20101 Nef variants and resembles the N\u2010terminal domain of NAP\u201022\/CAP23, a myristoylated calmodulin\u2010binder. These results for the interaction between HIV Nef and calmodulin in the cells suggested that the Nef might interfere with intracellular Ca<jats:sup>2+<\/jats:sup> signaling through calmodulin\u2010mediated interactions in infected cells.<\/jats:p>","DOI":"10.1110\/ps.04969605","type":"journal-article","created":{"date-parts":[[2005,1,5]],"date-time":"2005-01-05T02:43:19Z","timestamp":1104892999000},"page":"494-503","source":"Crossref","is-referenced-by-count":27,"title":["Myristoyl moiety of HIV Nef is involved in regulation of the interaction with calmodulin in vivo"],"prefix":"10.1002","volume":"14","author":[{"given":"Mamoru","family":"Matsubara","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Jing","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kumi","family":"Kawamura","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Naoshi","family":"Shimojo","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Koiti","family":"Titani","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keiichiro","family":"Hashimoto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nobuhiro","family":"Hayashi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2009,1]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0969-2126(97)00286-4"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi9629945"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1074-7613(00)80331-3"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.94.7.3178"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.11.6333"},{"key":"e_1_2_7_7_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.bb.24.060195.000505"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0960-9822(02)70770-7"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1089\/aid.1994.10.1231"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1089\/aid.1998.14.1543"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.1470917"},{"key":"e_1_2_7_12_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.270.5238.988"},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.4.1506"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1997.00843.x"},{"key":"e_1_2_7_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0264-410X(93)90049-4"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.69.7.4112-4121.1995"},{"key":"e_1_2_7_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/0006-291X(82)90712-4"},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.12.6590"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1994.1027"},{"key":"e_1_2_7_20_1","doi-asserted-by":"crossref","first-page":"6701","DOI":"10.1128\/jvi.70.10.6701-6708.1996","article-title":"Human immunodeficiency virus type 1 Nef binds directly to Lck and mitogen\u2010activated protein kinase, inhibiting kinase activity","volume":"70","author":"Greenway A.","year":"1996","journal-title":"J. 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