0 9 Efficient efficient JJ 10 23 transcription transcription NN 24 27 and and CC 28 39 replication replication NN 40 42 of of IN 43 49 simian simian JJ 50 66 immunodeficiency immunodeficiency NN 67 72 virus virus NN 73 75 in in IN 76 79 the the DT 80 87 absence absence NN 88 90 of of IN 91 100 NF-kappaB NF-kappaB NNP 101 104 and and CC 105 108 Sp1 sp1 NN 109 116 binding binding NN 117 125 elements element NNS 125 126 . . . 128 131 Ten ten CD 132 139 mutants mutant NNS 140 142 of of IN 143 146 the the DT 147 153 simian simian JJ 154 170 immunodeficiency immunodeficiency NN 171 176 virus virus NN 177 178 ( ( ( 178 181 SIV SIV NNP 181 182 ) ) ) 183 192 SIVmac239 SIVmac239 NNP 193 200 bearing bear VBG 201 210 deletions deletion NNS 211 212 ( ( ( 212 217 delta delta SYM 217 218 ) ) ) 219 221 or or CC 222 235 substitutions substitution NNS 236 237 ( ( ( 237 242 subst subst NN 242 243 ) ) ) 244 246 in in IN 247 250 the the DT 251 260 NF-kappaB NF-kappaB NNP 261 267 and/or and/or CC 268 271 Sp1 sp1 NN 272 279 binding binding NN 280 288 elements element NNS 289 293 were be VBD 294 301 created create VBN 301 302 , , , 303 306 and and CC 307 310 the the DT 311 322 replicative replicative JJ 323 333 capacities capacity NNS 334 336 of of IN 337 340 the the DT 341 348 mutants mutant NNS 349 353 were be VBD 354 362 analyzed analyze VBN 362 363 . . . 364 367 All all DT 368 375 mutants mutant NNS 375 376 , , , 377 386 including include VBG 387 390 one one CD 391 402 extensively extensively RB 403 414 mutagenized mutagenize VBN 415 421 strain strain NN 422 430 entirely entirely RB 431 438 missing miss VBG 439 442 the the DT 443 452 NF-kappaB NF-kappaB NNP 453 456 and and CC 457 461 four four CD 462 465 Spl spl NN 466 473 binding binding NN 474 482 elements element NNS 482 483 , , , 484 494 replicated replicate VBN 495 499 with with IN 500 509 wild-type wild-type JJ 510 518 kinetics kinetic NNS 519 522 and and CC 523 525 to to TO 526 527 a a DT 528 537 wild-type wild-type JJ 538 543 level level NN 544 546 in in IN 547 557 peripheral peripheral JJ 558 563 blood blood NN 564 575 mononuclear mononuclear JJ 576 580 cell cell NN 581 589 cultures culture NNS 590 592 in in IN 593 595 50 50 CD 596 598 to to TO 599 602 100 100 CD 602 603 % % NN 604 606 of of IN 607 610 the the DT 611 622 experiments experiment NNS 622 623 . . . 624 627 One one CD 628 633 group group NN 634 636 of of IN 637 644 mutants mutant NNS 645 655 replicated replicate VBN 656 660 very very RB 661 670 similarly similarly RB 671 673 to to TO 674 683 SIVmac239 SIVmac239 NNP 684 686 in in IN 687 695 kinetics kinetic NNS 696 699 and and CC 700 705 yield yield NN 706 708 in in IN 709 716 CEMxl74 cemxl74 NN 717 722 cells cell NNS 723 724 ( ( ( 724 734 2xNFKappaB 2xNFKappaB NNP 735 736 > > SYM 737 739 or or CC 740 741 = = SYM 742 751 SlVmac239 slvmac239 NN 752 765 approximately approximately RB 766 779 deltaNFkappaB deltanfkappab NN 780 793 approximately approximately RB 794 805 deltaSpl234 deltaspl234 NN 806 819 approximately approximately RB 820 833 substNFkappaB substnfkappab NN 834 847 approximately approximately RB 848 857 substSpl2 substspl2 NN 858 871 approximately approximately RB 872 881 substSp23 substsp23 NN 881 882 ) ) ) 882 883 , , , 884 889 while while IN 890 891 a a DT 892 898 second second JJ 899 904 group group NN 905 915 replicated replicate VBN 916 920 with with IN 921 928 delayed delay VBN 929 931 or or CC 932 940 slightly slightly RB 941 948 delayed delay VBN 949 957 kinetics kinetic NNS 958 960 in in IN 961 968 CEMxl74 cemxl74 NN 969 974 cells cell NNS 975 976 ( ( ( 976 985 SIVmac239 SIVmac239 NNP 986 987 > > SYM 988 997 substSp34 substsp34 NN 998 999 > > SYM 1000 1024 deltaNFkappaBdeltaSpl234 deltanfkappabdeltaspl234 NN 1025 1038 approximately approximately RB 1039 1060 deltaNFkappaBdeltaSp1 deltanfkappabdeltasp1 NN 1061 1062 > > JJ 1063 1074 substSpl234 substspl234 NN 1074 1075 ) ) ) 1075 1076 . . . 1077 1087 Reversions reversion NNS 1088 1090 or or CC 1091 1101 additional additional JJ 1102 1111 mutations mutation NNS 1112 1116 were be VBD 1117 1120 not not RB 1121 1129 detected detect VBN 1130 1132 in in IN 1133 1136 the the DT 1137 1139 U3 u3 NN 1140 1143 and and CC 1144 1145 R r NN 1146 1153 regions region NNS 1154 1156 of of IN 1157 1165 proviral proviral JJ 1166 1169 DNA dna NN 1170 1174 from from IN 1175 1182 CEMxl74 cemxl74 NN 1183 1188 cells cell NNS 1189 1197 infected infect VBN 1198 1202 with with IN 1203 1206 the the DT 1207 1216 SIVmac239 SIVmac239 NNP 1217 1224 mutants mutant NNS 1224 1225 . . . 1226 1233 Similar similar JJ 1234 1241 results result NNS 1242 1246 were be VBD 1247 1255 obtained obtain VBN 1256 1260 when when WRB 1261 1268 mutants mutant NNS 1269 1271 of of IN 1272 1281 SIVmacMER SIVmacMER NNP 1282 1283 ( ( ( 1283 1284 a a DT 1285 1305 macrophage-competent macrophage-competent JJ 1306 1316 derivative derivative NN 1317 1319 of of IN 1320 1329 SIVmac239 SIVmac239 NNP 1329 1330 ) ) ) 1331 1335 were be VBD 1336 1342 tested test VBN 1343 1345 in in IN 1346 1356 peripheral peripheral JJ 1357 1362 blood blood NN 1363 1374 mononuclear mononuclear JJ 1375 1379 cell cell NN 1380 1383 and and CC 1384 1391 CEMx174 cemx174 NN 1392 1400 cultures culture NNS 1400 1401 . . . 1402 1409 However however RB 1409 1410 , , , 1411 1414 the the DT 1415 1421 growth growth NN 1422 1424 of of IN 1425 1429 most most JJS 1430 1437 mutated mutate VBN 1438 1445 viruses virus NNS 1446 1449 was be VBD 1450 1460 suppressed suppress VBN 1461 1463 in in IN 1464 1471 primary primary JJ 1472 1478 rhesus rhesus NN 1479 1485 monkey monkey NN 1486 1494 alveolar alveolar JJ 1495 1506 macrophages macrophage NNS 1507 1508 ( ( ( 1508 1517 SIVmacMER SIVmacMER NNP 1518 1531 approximately approximately RB 1532 1542 2xNFkappaB 2xnfkappab NN 1543 1556 approximately approximately RB 1557 1570 substNFkappaB substnfkappab NN 1571 1572 > > SYM 1573 1586 deltaNFkappaB deltaNFkappaB NNP 1587 1588 > > SYM 1589 1613 deltaNFkappaBdeltaSpl234 deltanfkappabdeltaspl234 NN 1614 1627 approximately approximately RB 1628 1649 deltaNFkappaBdeltaSpl deltanfkappabdeltaspl NN 1650 1651 > > JJ 1652 1663 deltaSpl234 deltaspl234 NN 1664 1677 approximately approximately RB 1678 1687 substSpl2 substspl2 NN 1688 1689 > > JJ 1690 1699 substSp23 substsp23 NN 1700 1713 approximately approximately RB 1714 1723 substSp34 substsp34 NN 1724 1737 approximately approximately RB 1738 1749 substSpl234 substspl234 NN 1750 1751 > > JJ 1752 1754 or or CC 1755 1756 = = JJ 1757 1766 SIVmac239 SIVmac239 NNP 1766 1767 ) ) ) 1767 1768 . . . 1769 1773 Thus thus RB 1773 1774 , , , 1775 1782 changes change NNS 1783 1785 in in IN 1786 1789 the the DT 1790 1793 Sp1 sp1 NN 1794 1801 binding binding NN 1802 1807 sites site NNS 1808 1811 had have VBD 1812 1815 the the DT 1816 1820 most most RBS 1821 1829 dramatic dramatic JJ 1830 1837 effects effect NNS 1838 1840 on on IN 1841 1847 SIVmac SIVmac NNP 1848 1859 replication replication NN 1860 1862 in in IN 1863 1870 primary primary JJ 1871 1881 macrophage macrophage NN 1882 1890 cultures culture NNS 1890 1891 . . . 1892 1900 Analysis Analysis NNP 1901 1903 of of IN 1904 1908 long long JJ 1909 1917 terminal terminal JJ 1918 1931 repeat-driven repeat-driven JJ 1932 1940 secreted secrete VBN 1941 1949 alkaline alkaline NN 1950 1961 phosphatase phosphatase NN 1962 1970 activity activity NN 1971 1973 in in IN 1974 1983 transient transient JJ 1984 1990 assays assay NNS 1991 1997 showed show VBD 1998 2002 that that IN 2002 2003 , , , 2004 2010 unlike unlike IN 2011 2016 human human JJ 2017 2033 immunodeficiency immunodeficiency NN 2034 2039 virus virus NN 2040 2044 type type NN 2045 2046 1 1 CD 2046 2047 , , , 2048 2051 the the DT 2052 2055 SIV SIV NNP 2056 2060 long long JJ 2061 2069 terminal terminal JJ 2070 2076 repeat repeat NN 2077 2086 possesses possess VBZ 2087 2089 an an DT 2090 2098 enhancer enhancer NN 2099 2105 region region NN 2106 2110 just just RB 2111 2119 upstream upstream RB 2120 2122 of of IN 2123 2126 the the DT 2127 2136 NF-kappaB NF-kappaB NNP 2137 2144 element element NN 2145 2150 which which WDT 2151 2160 maintains maintain VBZ 2161 2172 significant significant JJ 2173 2179 levels level NNS 2180 2182 of of IN 2183 2188 basal basal JJ 2189 2202 transcription transcription NN 2203 2205 in in IN 2206 2209 the the DT 2210 2217 absence absence NN 2218 2220 of of IN 2221 2230 NF-kappaB NF-kappaB NNP 2231 2234 and and CC 2235 2238 Sp1 sp1 NN 2239 2244 sites site NNS 2244 2245 . . . 2246 2250 This this DT 2251 2257 region region NN 2258 2260 is be VBZ 2261 2271 responsive responsive JJ 2272 2274 to to TO 2275 2290 transactivation transactivation NN 2291 2293 by by IN 2294 2297 Tat Tat NNP 2297 2298 . . . 2299 2301 In in IN 2302 2310 addition addition NN 2310 2311 , , , 2312 2315 the the DT 2316 2319 SIV SIV NNP 2320 2324 TATA TATA NNP 2325 2328 box box NN 2329 2332 was be VBD 2333 2338 shown show VBN 2339 2341 to to TO 2342 2344 be be VB 2345 2353 stronger strong JJR 2354 2358 than than IN 2359 2363 that that DT 2364 2366 of of IN 2367 2372 human human JJ 2373 2389 immunodeficiency immunodeficiency NN 2390 2395 virus virus NN 2396 2400 type type NN 2401 2402 1 1 CD 2402 2403 . . . 2404 2413 Therefore therefore RB 2413 2414 , , , 2415 2418 the the DT 2419 2431 surprisingly surprisingly RB 2432 2436 high high JJ 2437 2448 replicative replicative JJ 2449 2457 capacity capacity NN 2458 2460 of of IN 2461 2470 NF-kappaB NF-kappaB NNP 2471 2474 and and CC 2475 2478 Sp1 sp1 NN 2479 2486 binding binding NN 2487 2491 site site NN 2492 2499 mutants mutant NNS 2500 2502 of of IN 2503 2509 SIVmac SIVmac NNP 2510 2512 is be VBZ 2513 2516 due due JJ 2517 2519 to to TO 2520 2526 unique unique JJ 2527 2535 features feature NNS 2536 2538 or or CC 2539 2542 the the DT 2543 2560 enhancer/promoter enhancer/promoter NN 2561 2567 region region NN 2567 2568 . . .