Characterization of human T-cell leukemia virus type I integrase expressed in Escherichia coli.

Characterization of human T-cell leukemia virus type I integrase expressed in Escherichia coli.
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在大肠杆菌中表达的人 T 细胞白血病病毒 I 型整合酶的表征。

DOI:
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发表时间:
1999
期刊:
European Journal of Biochemistry
影响因子:
--
通讯作者:
H. Kräusslich
H. Kräusslich
中科院分区:
--
文献类型:
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作者:
B. Müller;H. Kräusslich

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预测人t细胞白血病病毒I型(HTLV-I) pol基因c端编码病毒整合酶(IN);然而,这种蛋白尚未在病毒粒子或感染细胞中检测到。我们在大肠杆菌中表达了HTLV-I传染性分子克隆的推定IN。与HTLV-I蛋白酶(PR)和Pol在昆虫细胞中共表达产生的蛋白比较表明,细菌表达的蛋白与PR-Pol前体通过蛋白水解裂解释放的in相同或非常相似。HTLV-I IN在天然条件下从大肠杆菌中纯化。该蛋白在尺寸排除层析中表现为二聚体。它高效地发挥逆转录病毒IN的活性特征,在加工和链转移反应中对u5衍生序列和u3衍生序列表现出强烈的偏好。在分解反应中,HTLV-I In不仅接受链转移反应产物对应的双链支链底物,而且能够在单链或单腺苷悬垂的支链分子上进行磷酸化转移。
The C-terminal part of the pol gene of the human T-cell leukemia virus type I (HTLV-I) is predicted to encode the integrase (IN) of the virus; however, this protein has not yet been detected in virions or infected cells. We expressed the putative IN from an infectious molecular clone of HTLV-I in Escherichia coli. Comparison with protein resulting from coexpression of HTLV-I protease (PR) and Pol in insect cells indicated that the bacterially expressed protein is identical with or very similar to IN released from a PR-Pol precursor by proteolytic cleavage. HTLV-I IN was purified from E. coli under native conditions. The protein behaved like a dimer in size-exclusion chromatography. It carried out activities characteristic of retroviral IN with high efficiency, displaying a strong preference for U5-derived vs. U3-derived sequences in the processing and strand-transfer reactions. In the disintegration reaction, HTLV-I IN not only accepted the double-stranded branched substrate corresponding to the product of a strand-transfer reaction, but was also able to carry out a phosphoryl transfer on a branched molecule with a single-stranded or a single adenosine overhang.
逆转录病毒整合酶以多聚体的形式发挥作用,并且可以催化翻转。
DOI: --
发表时间: 1992
期刊: The Journal of biological chemistry
影响因子: --
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人类 T 细胞白血病病毒 II 型整合酶的催化活性。
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