Activity of HIV-1 integrases recovered from subjects with varied rates of disease progression.
Activity of HIV-1 integrases recovered from subjects with varied rates of disease progression.
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从疾病进展速度不同的受试者中恢复了 HIV-1 整合酶的活性。
DOI：
10.1097/00042560-200111010-00001
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Eyster,ME
中科院分区：
文献类型：
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作者：
Katzman,M;Harper,AL;Sudol,M;Skinner,LM;Eyster,ME
我们最近描述了102个HIV-1整合酶序列，这些序列是从18年前从5名后来死于艾滋病的血友病患者和5名后来被归类为缓慢或非进展的血友病患者的血细胞或血浆中扩增出来的（J acquired Immune Defic Syndr Hum Retrovirol 1998; 19: 99-110）。尽管HIV-1基因组编码整合酶的区域是高度保守的，但推导出的患者衍生酶的蛋白质序列都与B支共识或标准实验室整合酶的蛋白质序列不匹配。为了验证HIV-1整合酶在感染者体内普遍存在的活性与疾病进展速度有关的假设，我们现在表达和纯化了这些蛋白质，并在各种检测中对它们进行了比较。从102个克隆中分离得到75个独特的全长整合酶蛋白，其中大部分具有酶活性。从感染后不久获得的样品中获得的蛋白质的比较表明，病毒DNA加工的特异性和程度以及DNA连接的数量（整合酶的两种生物学相关活性）在两组患者之间没有差异。此外，替代亲核试剂的相对使用和由蛋白质催化的非特异性缺口的数量在患者组之间是无法区分的。尽管与实验室整合酶相比，患者源性酶通常表现出不同的靶位偏好模式，但与临床病程无关。因此，在这些感染者中普遍存在的HIV-1整合酶的活性，至少在标准测定中反映出来，并不影响或预测疾病进展的速度。
: We recently described 102 HIV-1 integrase sequences that were amplified from blood cells or plasma obtained up to 18 years ago from 5 hemophiliacs who later died of AIDS and 5 hemophiliacs subsequently classified as slow or nonprogressors (J Acquir Immune Defic Syndr Hum Retrovirol 1998; 19: 99-110). Although the region of the HIV-1 genome that encodes integrase was highly conserved, none of the deduced protein sequences of the patient-derived enzymes matched that of the clade B consensus or standard laboratory integrases. To test the hypothesis that the activity of HIV-1 integrases prevalent within an infected person contributes to the rate of disease progression, we have now expressed and purified these proteins and compared them in various assays. Most of the 75 unique full-length integrase proteins from the 102 clones were enzymatically active. Comparison of proteins derived from samples obtained soon after infection showed that the specificity and extent of viral DNA processing and the amount of DNA joining (the two biologically relevant activities of integrase) did not differ between the two groups of patients. In addition, the relative usage of alternative nucleophiles for processing and the amount of nonspecific nicking catalyzed by the proteins were indistinguishable between the patient groups. Although the patient-derived enzymes often exhibited different patterns of target site preferences compared with the laboratory integrase, there was no correlation with clinical course. Thus, the activities of HIV-1 integrases prevalent within these infected individuals, at least as reflected by standard assays, did not influence or predict the rate of disease progression.