0 1 A a DT 2 10 mutation mutation NN 11 13 of of IN 14 17 the the DT 18 32 glucocorticoid glucocorticoid NN 33 41 receptor receptor NN 42 44 in in IN 45 52 primary primary JJ 53 61 cortisol cortisol NN 62 72 resistance resistance NN 72 73 . . . 75 78 The the DT 79 86 precise precise JJ 87 96 molecular molecular JJ 97 110 abnormalities abnormality NNS 111 115 that that WDT 116 121 cause cause VBP 122 129 primary primary JJ 130 138 cortisol cortisol NN 139 149 resistance resistance NN 150 154 have have VBP 155 158 not not RB 159 163 been be VBN 164 174 completely completely RB 175 184 described describe VBN 184 185 . . . 186 188 In in IN 189 190 a a DT 191 198 subject subject NN 199 203 with with IN 204 211 primary primary JJ 212 220 cortisol cortisol NN 221 231 resistance resistance NN 232 234 we we PRP 235 239 have have VBP 240 248 observed observe VBN 249 263 glucocorticoid glucocorticoid NN 264 273 receptors receptor NNS 274 275 ( ( ( 275 278 hGR hGR NNP 278 279 ) ) ) 280 284 with with IN 285 286 a a DT 287 296 decreased decrease VBN 297 305 affinity affinity NN 306 309 for for IN 310 323 dexamethasone dexamethasone NN 323 324 . . . 325 327 We we PRP 328 339 hypothesize hypothesize VBP 340 344 that that IN 345 346 a a DT 347 355 mutation mutation NN 356 358 of of IN 359 362 the the DT 363 366 hGR hGR NNP 367 389 glucocorticoid-binding glucocorticoid-binding JJ 390 396 domain domain NN 397 399 is be VBZ 400 403 the the DT 404 409 cause cause NN 410 412 of of IN 413 421 cortisol cortisol NN 422 432 resistance resistance NN 432 433 . . . 434 439 Total total JJ 440 443 RNA RNA NNP 444 452 isolated isolate VBN 453 457 from from IN 458 461 the the DT 462 467 index index NN 468 475 subject subject NN 475 477 ’s 's POS 478 489 mononuclear mononuclear JJ 490 500 leukocytes leukocyte NNS 501 504 was be VBD 505 509 used use VBN 510 512 to to TO 513 520 produce produce VB 521 526 first first JJ 527 533 strand strand NN 534 537 hGR hGR NNP 538 543 cDNAs cdna NNS 543 544 , , , 545 548 and and CC 549 552 the the DT 553 559 entire entire JJ 560 563 hGR hGR NNP 564 568 cDNA cDNA NNP 569 572 was be VBD 573 582 amplified amplify VBN 583 585 in in IN 586 594 segments segment NNS 595 598 and and CC 599 608 sequenced sequence VBN 608 609 . . . 610 612 At at IN 613 623 nucleotide nucleotide NN 624 629 2,317 2,317 CD 630 632 we we PRP 633 643 identified identify VBD 644 645 a a DT 646 656 homozygous homozygous JJ 657 658 A a NN 659 662 for for IN 663 664 G G NNP 665 670 point point NN 671 679 mutation mutation NN 680 684 that that WDT 685 693 predicts predict VBZ 694 696 an an DT 697 707 isoleucine isoleucine NN 708 709 ( ( ( 709 712 ATT ATT NNP 712 713 ) ) ) 714 717 for for IN 718 724 valine valine NN 725 726 ( ( ( 726 729 GTT GTT NNP 729 730 ) ) ) 731 743 substitution substitution NN 744 746 at at IN 747 752 amino amino NN 753 757 acid acid NN 758 761 729 729 CD 761 762 . . . 763 767 When when WRB 768 771 the the DT 772 781 wild-type wild-type JJ 782 785 hGR hGR NNP 786 789 and and CC 790 797 hGR-Ile hgr-ile NN 798 801 729 729 CD 802 806 were be VBD 807 816 expressed express VBN 817 819 in in IN 820 825 COS-1 cos-1 NN 826 831 cells cell NNS 832 835 and and CC 836 843 assayed assay VBN 844 847 for for IN 848 849 [ [ ( 849 866 3H]-Dexamethasone 3H]-Dexamethasone NNP 867 874 binding binding NN 874 875 , , , 876 879 the the DT 880 892 dissociation dissociation NN 893 902 constants constant NNS 903 907 were be VBD 908 913 0.799 0.799 CD 914 917 +/- +/- SYM 918 923 0.068 0.068 CD 924 927 and and CC 928 932 1.54 1.54 CD 933 936 +/- +/- SYM 937 941 0.06 0.06 CD 942 944 nM nm NN 945 946 ( ( ( 946 950 mean mean NN 951 954 +/- +/- CC 955 958 SEM SEM NNP 958 959 ) ) ) 960 961 ( ( ( 961 962 P p NN 963 964 < < SYM 965 969 0.01 0.01 CD 969 970 ) ) ) 970 971 , , , 972 984 respectively respectively RB 984 985 . . . 986 990 When when WRB 991 994 the the DT 995 1004 wild-type wild-type JJ 1005 1008 hGR hGR NNP 1009 1012 and and CC 1013 1020 hGR-Ile hgr-ile NN 1021 1024 729 729 CD 1025 1029 were be VBD 1030 1039 expressed express VBN 1040 1042 in in IN 1043 1047 CV-1 cv-1 NN 1048 1053 cells cell NNS 1054 1058 that that WDT 1059 1063 were be VBD 1064 1077 cotransfected cotransfecte VBN 1078 1082 with with IN 1083 1086 the the DT 1087 1092 mouse mouse NN 1093 1100 mammary mammary JJ 1101 1106 tumor tumor NN 1107 1112 virus virus NN 1113 1117 long long JJ 1118 1126 terminal terminal JJ 1127 1133 repeat repeat NN 1134 1139 fused fuse VBN 1140 1142 to to TO 1143 1146 the the DT 1147 1162 chloramphenicol chloramphenicol NN 1163 1169 acetyl acetyl NN 1170 1181 transferase transferase NN 1182 1183 ( ( ( 1183 1186 CAT CAT NNP 1186 1187 ) ) ) 1188 1192 gene gene NN 1192 1193 , , , 1194 1197 the the DT 1198 1205 hGR-Ile hgr-ile NN 1206 1209 729 729 CD 1210 1219 conferred confer VBD 1220 1221 a a DT 1222 1230 fourfold fourfold JJ 1231 1239 decrease decrease NN 1240 1242 in in IN 1243 1251 apparent apparent JJ 1252 1259 potency potency NN 1260 1262 on on IN 1263 1276 dexamethasone dexamethasone NN 1277 1288 stimulation stimulation NN 1289 1291 of of IN 1292 1295 CAT CAT NNP 1296 1304 activity activity NN 1304 1305 . . . 1306 1309 The the DT 1310 1320 isoleucine isoleucine NN 1321 1324 for for IN 1325 1331 valine valine NN 1332 1344 substitution substitution NN 1345 1347 at at IN 1348 1353 amino amino NN 1354 1358 acid acid NN 1359 1362 729 729 CD 1363 1370 impairs impair VBZ 1371 1374 the the DT 1375 1383 function function NN 1384 1386 of of IN 1387 1390 the the DT 1391 1394 hGR hGR NNP 1395 1398 and and CC 1399 1401 is be VBZ 1402 1405 the the DT 1406 1412 likely likely JJ 1413 1418 cause cause NN 1419 1421 of of IN 1422 1429 primary primary JJ 1430 1438 cortisol cortisol NN 1439 1449 resistance resistance NN 1450 1452 in in IN 1453 1457 this this DT 1458 1465 subject subject NN 1465 1466 . . .