0 3 The the DT 4 15 involvement involvement NN 16 18 of of IN 19 27 multiple multiple JJ 28 33 tumor tumor NN 34 42 necrosis necrosis NN 43 49 factor factor NN 50 58 receptor receptor NN 59 60 ( ( ( 60 76 TNFR)-associated tnfr)-associated JJ 77 84 factors factor NNS 85 87 in in IN 88 91 the the DT 92 101 signaling signaling NN 102 112 mechanisms mechanism NNS 113 115 of of IN 116 124 receptor receptor NN 125 134 activator activator NN 135 137 of of IN 138 147 NF-kappaB NF-kappaB NNP 147 148 , , , 149 150 a a DT 151 157 member member NN 158 160 of of IN 161 164 the the DT 165 169 TNFR tnfr NN 170 181 superfamily superfamily NN 181 182 . . . 184 192 Receptor receptor NN 193 202 activator activator NN 203 205 of of IN 206 215 NF-kappaB NF-kappaB NNP 216 217 ( ( ( 217 221 RANK RANK NNP 221 222 ) ) ) 223 225 is be VBZ 226 227 a a DT 228 236 recently recently RB 237 247 identified identify VBN 248 254 member member NN 255 257 of of IN 258 261 the the DT 262 267 tumor tumor NN 268 276 necrosis necrosis NN 277 283 factor factor NN 284 292 receptor receptor NN 293 304 superfamily superfamily NN 305 308 and and CC 309 311 is be VBZ 312 321 expressed express VBN 322 324 on on IN 325 334 activated activate VBN 335 336 T t NN 337 342 cells cell NNS 343 346 and and CC 347 356 dendritic dendritic JJ 357 362 cells cell NNS 362 363 . . . 364 367 Its its PRP$ 368 375 cognate cognate JJ 376 382 ligand ligand NN 383 384 ( ( ( 384 389 RANKL RANKL NNP 389 390 ) ) ) 391 396 plays play VBZ 397 408 significant significant JJ 409 414 roles role NNS 415 417 in in IN 418 421 the the DT 422 432 activation activation NN 433 435 of of IN 436 445 dendritic dendritic JJ 446 450 cell cell NN 451 459 function function NN 460 463 and and CC 464 474 osteoclast osteoclast NN 475 490 differentiation differentiation NN 490 491 . . . 492 494 We we PRP 495 506 demonstrate demonstrate VBP 507 511 here here RB 512 515 the the DT 516 527 interaction interaction NN 528 530 of of IN 531 535 RANK RANK NNP 536 540 with with IN 541 546 tumor tumor NN 547 555 necrosis necrosis NN 556 562 factor factor NN 563 582 receptor-associated receptor-associated JJ 583 590 factors factor NNS 591 592 ( ( ( 592 597 TRAFs TRAF NNP 597 598 ) ) ) 599 600 1 1 CD 600 601 , , , 602 603 2 2 CD 603 604 , , , 605 606 3 3 CD 606 607 , , , 608 609 5 5 CD 609 610 , , , 611 614 and and CC 615 616 6 6 CD 617 621 both both CC 622 624 in in FW 625 630 vitro vitro FW 631 634 and and CC 635 637 in in IN 638 643 cells cell NNS 643 644 . . . 645 652 Mapping mapping NN 653 655 of of IN 656 659 the the DT 660 670 structural structural JJ 671 683 requirements requirement NNS 684 687 for for IN 688 697 TRAF/RANK TRAF/RANK NNP 698 709 interaction interaction NN 710 718 revealed reveal VBD 719 727 multiple multiple JJ 728 732 TRAF traf NN 733 740 binding binding NN 741 746 sites site NNS 747 756 clustered cluster VBN 757 759 in in IN 760 763 two two CD 764 772 distinct distinct JJ 773 780 domains domain NNS 781 783 in in IN 784 787 the the DT 788 792 RANK RANK NNP 793 804 cytoplasmic cytoplasmic JJ 805 809 tail tail NN 809 810 . . . 811 816 These these DT 817 821 TRAF TRAF NNP 822 829 binding binding NN 830 837 domains domain NNS 838 842 were be VBD 843 848 shown show VBN 849 851 to to TO 852 854 be be VB 855 867 functionally functionally RB 868 877 important important JJ 878 881 for for IN 882 885 the the DT 886 900 RANK-dependent rank-dependent JJ 901 910 induction induction NN 911 913 of of IN 914 923 NF-kappaB NF-kappaB NNP 924 927 and and CC 928 933 c-Jun c-jun NN 934 946 NH2-terminal nh2-terminal JJ 947 953 kinase kinase NNP 954 964 activities activity NNS 964 965 . . . 966 979 Site-directed site-directed JJ 980 991 mutagenesis mutagenesis NN 992 1004 demonstrated demonstrate VBD 1005 1009 that that IN 1010 1015 these these DT 1016 1020 TRAF TRAF NNP 1021 1028 binding binding NN 1029 1034 sites site NNS 1035 1044 exhibited exhibit VBD 1045 1054 selective selective JJ 1055 1062 binding binding NN 1063 1066 for for IN 1067 1076 different different JJ 1077 1081 TRAF TRAF NNP 1082 1090 proteins protein NNS 1090 1091 . . . 1092 1094 In in IN 1095 1105 particular particular JJ 1105 1106 , , , 1107 1112 TRAF6 TRAF6 NNP 1113 1123 interacted interact VBD 1124 1128 with with IN 1129 1146 membrane-proximal membrane-proximal JJ 1147 1159 determinants determinant NNS 1160 1168 distinct distinct JJ 1169 1173 from from IN 1174 1179 those those DT 1180 1187 binding bind VBG 1188 1193 TRAFs TRAF NNP 1194 1195 1 1 CD 1195 1196 , , , 1197 1198 2 2 CD 1198 1199 , , , 1200 1201 3 3 CD 1201 1202 , , , 1203 1206 and and CC 1207 1208 5 5 CD 1208 1209 . . . 1210 1214 When when WRB 1215 1219 this this DT 1220 1237 membrane-proximal membrane-proximal JJ 1238 1243 TRAF6 traf6 NN 1244 1255 interaction interaction NN 1256 1262 domain domain NN 1263 1266 was be VBD 1267 1274 deleted delete VBN 1274 1275 , , , 1276 1289 RANK-mediated rank-mediated JJ 1290 1299 NF-kappaB NF-kappaB NNP 1300 1309 signaling signaling NN 1310 1313 was be VBD 1314 1324 completely completely RB 1325 1334 inhibited inhibit VBN 1335 1340 while while IN 1341 1346 c-Jun c-jun NN 1347 1359 NH2-terminal nh2-terminal JJ 1360 1366 kinase kinase NNP 1367 1377 activation activation NN 1378 1381 was be VBD 1382 1391 partially partially RB 1392 1401 inhibited inhibit VBN 1401 1402 . . . 1403 1405 An an DT 1406 1418 NH2-terminal nh2-terminal JJ 1419 1429 truncation truncation NN 1430 1436 mutant mutant NN 1437 1439 of of IN 1440 1445 TRAF6 TRAF6 NNP 1446 1455 inhibited inhibit VBD 1456 1470 RANKL-mediated rankl-mediated JJ 1471 1480 NF-kappaB NF-kappaB NNP 1481 1491 activation activation NN 1491 1492 , , , 1493 1496 but but CC 1497 1503 failed fail VBD 1504 1506 to to TO 1507 1513 affect affect VB 1514 1526 constitutive constitutive JJ 1527 1536 signaling signaling NN 1537 1544 induced induce VBN 1545 1547 by by IN 1548 1556 receptor receptor NN 1557 1571 overexpression overexpression NN 1571 1572 , , , 1573 1582 revealing reveal VBG 1583 1584 a a DT 1585 1594 selective selective JJ 1595 1599 role role NN 1600 1603 for for IN 1604 1609 TRAF6 traf6 NN 1610 1612 in in IN 1613 1627 ligand-induced ligand-induced JJ 1628 1638 activation activation NN 1639 1645 events event NNS 1645 1646 . . .