The HIV-1 integrase monomer induces a specific interaction with LTR DNA for concerted integration.
The HIV-1 integrase monomer induces a specific interaction with LTR DNA for concerted integration.
复制标题
HIV-1整合酶单体诱导与LTR DNA的特定相互作用进行协同整合。
DOI：
10.1021/bi201247f
复制
发表时间：
2011-11-15
期刊：
影响因子：
2.9
通讯作者：
Grandgenett, Duane P.
中科院分区：
文献类型：
--
作者：
Pandey, Krishan K.;Bera, Sibes;Grandgenett, Duane P.
人类免疫缺陷病毒1型（HIV）突触复合物（SC）能够协同整合的组装机制尚不清楚。分子和结构研究已经确定，HIV SC和原型泡沫病毒（PFV）整合体含有催化协同整合的整合酶（IN）四聚体。在1 mM EDTA和10 mM MgSO 4存在下纯化的HIV IN主要是单体。IN可有效地促进微摩尔浓度的3′ OH凹陷和平端U 5长末端重复序列（LTR）寡核苷酸（ODN）底物（19 bp ~ 42 bp）协同整合到环状靶DNA中。改变HIV IN与U 5 DNA显示IN二聚体：DNA末端摩尔比为1对于协同整合是最佳的。整合活性随着ODN长度的增加而降低，从凹陷的18/20 bp或19/21 bp组到31/33 bp和40/42 bp组。在这些条件下，用凹陷和平端底物进行HIV 5 bp宿主位点重复的平均保真度为56%。对较长DNA（1.6 kb）末端超过21 bp的U 5 LTR序列进行修饰不会改变~32 bp的DNaseI保护足迹，这表明超过21 bp的病毒序列对于IN结合不是必需的。结果表明，IN与18/20 bp的ODN底物的结合比与40/42 bp的ODN底物的结合差异更大。HIV IN单体可能是在不存在或存在链转移抑制剂的情况下尝试IN-DNA复合物结晶的合适候选物。
The assembly mechanism for the human immunodeficiency virus type-1 (HIV) synaptic complex (SC) capable of concerted integration is unknown. Molecular and structural studies have established that the HIV SC and prototype foamy virus (PFV) intasome contain a tetramer of integrase (IN) that catalyzes concerted integration. HIV IN purified in the presence of 1 mM EDTA and 10 mM MgSO4 was predominately a monomer. IN efficiently promoted concerted integration of micromolar concentrations of 3′ OH recessed and blunt ended U5 long terminal repeat (LTR) oligonucleotides (ODN) substrates (19 bp to 42 bp) into circular target DNA. Varying HIV IN to U5 DNA showed that a IN dimer:DNA end molar ratio of 1 was optimal for concerted integration. Integration activities decreased with increasing length of the ODN, starting from the recessed 18/20 bp or 19/21 bp set to the 31/33 bp and 40/42 bp set. Under these conditions, the average fidelity for the HIV 5 bp host site duplication with recessed and blunt ended substrates was 56%. Modifications of U5 LTR sequences beyond 21 bp from the terminus on longer DNA (1.6 kb) did not alter the ~32 bp DNaseI protective footprint suggesting, viral sequences beyond 21 bp were not essential for IN binding. The results suggest IN binds differentially to an 18/20 bp than to an 40/42 bp ODN substrate for concerted integration. The HIV IN monomer may be a suitable candidate to attempt crystallization of an IN-DNA complex in the absence or presence of strand transfer inhibitors.
登录
查看更多内容
新型的共晶结构为在经过改进的PSIP1/LEDGF/P75的存在下提供了功能获得的慢病毒积分突变体的设计。
影响因子：
6.7
作者：
Hare S;Shun MC;Gupta SS;Valkov E;Engelman A;Cherepanov P
通讯作者：
Cherepanov P
影响因子：
4.8
作者：
Jenkins, TM;Engelman, A;Craigie, R
通讯作者：
Craigie, R
DOI：
10.1073/pnas.87.11.4164
发表时间：
1990-06-01
影响因子：
11.1
作者：
FARNET, CM;HASELTINE, WA
通讯作者：
HASELTINE, WA
影响因子：
2.9
作者：
Deprez, E;Tauc, P;Brochon, JC
通讯作者：
Brochon, JC
影响因子：
2.9
作者：
Lee, SP;Xiao, JM;Han, MK
通讯作者：
Han, MK