POTENT INHIBITION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 (HIV-1) REPLICATION BY INDUCIBLE EXPRESSION OF HIV-1 PR MULTIMERS
POTENT INHIBITION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 (HIV-1) REPLICATION BY INDUCIBLE EXPRESSION OF HIV-1 PR MULTIMERS
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DOI：
10.1128/jvi.69.10.5988-5994.1995
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发表时间：
1995-10-01
影响因子：
5.4
通讯作者：
HUFFMAN, K
中科院分区：
文献类型：
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作者：
ARRIGO, SJ;HUFFMAN, K
产生构建体，其中人免疫缺陷病毒1型（HIV-1）蛋白酶（PR）的表达置于HIV-1长末端重复序列的控制下，因此需要HIV-1达特蛋白用于PR的表达。通过与产生Gag底物的构建体共转染来评估PR的活性。PR作为分子内多聚体的表达导致PR活性与PR作为单体的表达所获得的水平相比大幅增加。表达PR多聚体的构建体也表现出PR表达的细胞毒性作用。用产生PR作为连接的四聚体的构建体产生CD 4（+）T细胞系，并筛选PR活性和诱导性。在这些细胞系中的HIV-1的复制是几个数量级减少相比，在不表达PR的细胞系。在这些细胞系中的感染可以检测到感染后早期，但随着时间的推移消失。在感染细胞后，通过加入PR的特异性抑制剂U 75875（Upjohn），PR表达细胞系的感染可增加几个数量级，表明这些细胞中HIV-1复制的有效抑制直接归因于PR的表达。
Constructs were generated in which expression of human immunodeficiency virus type 1 (HIV-1) protease (PR) was placed under control of the HIV-1 long terminal repeat, thus requiring the HIV-1 Tat protein for expression of PR. The activity of PR was assessed by cotransfection with a construct producing a Gag substrate. Expression of PR as an intramolecular multimer resulted in a large increase in PR activity in comparison with the level obtained with the expression of PR as a monomer. A cytotoxic effect of PR expression was also exhibited by the constructs expressing PR multimers. CD4(+) T-cell lines were generated with a construct producing PR as a linked tetramer and screened for PR activity and inducibility. The replication of HIV-1 in these cell lines was several orders of magnitude reduced in comparison with that in cell lines not expressing PR. Infection in these cell lines could be detected early after infection but disappeared over time. Infection of the PR-expressing cell lines could be increased several orders of magnitude by the addition of a specific inhibitor of PR, U75875 (Upjohn), after infection of the cells, demonstrating that the potent inhibition of HIV-1 replication in these cells was directly due to the expression of PR.