COMPLETE MUTAGENESIS OF THE HIV-1 PROTEASE
COMPLETE MUTAGENESIS OF THE HIV-1 PROTEASE
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DOI：
10.1038/340397a0
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发表时间：
1989-08-03
期刊：
影响因子：
64.8
通讯作者：
HUTCHISON, CA
中科院分区：
文献类型：
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作者：
LOEB, DD;SWANSTROM, R;HUTCHISON, CA
逆转录病毒编码一种蛋白酶，这种蛋白酶需要有活性才能产生感染性病毒体。蛋白酶中的失能突变导致产生非感染性病毒颗粒，并且对来自这些突变病毒体的蛋白质的检查揭示了未加工的Gag和Gag-Pol前体蛋白，即病毒蛋白酶1 -3的底物。使用能够在蛋白质的每个残基中引入和鉴定错义突变的简单诱变程序单独地突变HIV-1蛋白酶的每个氨基酸。在异源测定系统中对这些突变体的表型筛选揭示了蛋白酶内的三个区域，其中多个连续的氨基酸残基对突变敏感。这些结果表明，随机诱变可用于鉴定蛋白质内的功能重要区域。具有条件表型的突变体也已在该集合中鉴定。
RETROVIRUSES encode a protease which needs to be active for the production of infectious virions. A disabling mutation in the protease results in the production of non-infectious virus particles and examination of proteins from these mutant virions reveals unprocessed Gag and Gag-Pol precursor proteins, the substrates of the viral protease1-3. Each amino acid of the HIV-1 protease was individually mutated using a simple mutagenesis procedure which is capable of introducing and identifying missense mutations in each residue of a protein. Phenotypic screening of these mutants in a heterologous assay system reveals three regions within the protease where multiple consecutive amino-acid residues are sensitive to mutation. These results show that random mutagenesis can be used to identify functionally important regions within a protein. Mutants with conditional phenotypes have also been identified within this collection.