RETROVIRAL INTEGRASE DOMAINS - DNA-BINDING AND THE RECOGNITION OF LTR SEQUENCES
RETROVIRAL INTEGRASE DOMAINS - DNA-BINDING AND THE RECOGNITION OF LTR SEQUENCES
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DOI：
10.1093/nar/19.4.851
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发表时间：
1991-02-25
影响因子：
14.9
通讯作者：
SKALKA, AM
中科院分区：
文献类型：
--
作者：
KHAN, E;MACK, JPG;SKALKA, AM
逆转录病毒DNA整合到宿主染色体需要病毒编码的整合酶(IN)。IN识别、切割并将特定的病毒DNA序列(LTR端)连接到宿主DNA中基本上随机的位置。我们使用了计算机辅助的蛋白质比对和突变，试图在禽类逆转录病毒IN蛋白中定位这些功能。对80个逆转录病毒/逆转录转座子IN蛋白推导的氨基酸序列进行比较，发现HHCCN端的锌指结构域和中心的D(35)E区具有很强的保守性，与细菌IS元件的推导序列具有显著的相似性。我们证明HHCC区域不是DNA结合所必需的，但在切割和连接反应中有助于病毒LTRs的特异性识别。延伸到D(35)E区的缺失破坏了IN结合DNA的能力。因此，我们认为D(35)E区可能指定了一个DNA结合/切割结构域，该结构域在具有相似功能的酶的整个进化过程中都是保守的。
Integration of retroviral DNA into the host chromosome requires a virus-encoded integrase (IN). IN recognizes, cuts and then joins specific viral DNA sequences (LTR ends) to essentially random sites in host DNA. We have used computer-assisted protein alignments and mutagenesis in an attempt to localize these functions within the avian retroviral IN protein. A comparison of the deduced amino acid sequences for 80 retroviral/retrotransposon IN proteins reveals strong conservation of an HHCC N-terminal 'Zn finger'-like domain, and a central D(35)E region which exhibits striking similarities with sequences deduced for bacterial IS elements. We demonstrate that the HHCC region is not required for DNA binding, but contributes to specific recognition of viral LTRs in the cutting and joining reactions. Deletions which extend into the D(35)E region destroy the ability of IN to bind DNA. Thus, we propose that the D(35)E region may specify a DNA-binding/cutting domain that is conserved throughout evolution in enzymes with similar functions.