Mutations in the catalytic core or the C-terminus of murine leukemia virus (MLV) integrase disrupt virion infectivity and exert diverse effects on reverse transcription

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• 作者：Adolf Steinrigl ; Dagmara Nosek ; Reinhard Ertl ; Walter H. Gü ; nzburg ; Brian Salmons ; Dieter Klein
• 关键词：MLV ; Integrase mutants ; Reverse transcription ; 2-LTR circles ; Real-time PCR
• 刊名：Virology
• 出版年：2007
• 出版时间：25 May 2007
• 年：2007
• 卷：362
• 期：1
• 页码：50-59
• 全文大小：331 K

文摘

Understanding of the structures and functions of the retroviral integrase (IN), a key enzyme in the viral replication cycle, is essential for developing antiretroviral treatments and facilitating the development of safer gene therapy vehicles. Thus, four MLV IN-mutants were constructed in the context of a retroviral vector system, harbouring either a substitution in the catalytic centre, deletions in the C-terminus, or combinations of both modifications. IN-mutants were tested for their performance in different stages of the viral replication cycle: RNA-packaging; RT-activity; transient and stable infection efficiency; dynamics of reverse transcription and nuclear entry. All mutant vectors packaged viral RNA with wild-type efficiencies and displayed only slight reductions in RT-activity. Deletion of either the IN C-terminus alone, or in addition to part of the catalytic domain exerted contrasting effects on intracellular viral DNA levels, implying that IN influences reverse transcription in more than one direction.