Escoubas J M, Lane D, Chandler M
Molecular Genetics and Microbiology, UPR9007 du Centre National de la Recherche Scientifique, Toulouse, France.
J Bacteriol. 1994 Sep;176(18):5864-7. doi: 10.1128/jb.176.18.5864-5867.1994.
The transposase of the bacterial insertion sequence IS1 is normally expressed by inefficient translational frameshifting between an upstream reading frame which itself specifies a transposition inhibitor, InsA, and a second consecutive reading frame located immediately downstream. A fused-frame mutant which carries an additional base pair inserted at the point of frameshifting was constructed. This mutant exhibits high transposition activity and should express the transposase, InsAB', constitutively without frameshifting. Unexpectedly, a second protein species was observed to be expressed from this mutant. We demonstrate here that this protein, InsA*, results from continued frameshifting on the modified frameshift motif. The protein retains the activities of the repressor InsA. Its elimination, by further modification of the frameshift motif, results in a further increase in various transposition activities of IS1. These results support the hypothesis that a single IS1-encoded protein, InsAB', is necessary for transposition.
细菌插入序列IS1的转座酶通常通过上游阅读框(其本身编码一种转座抑制剂InsA)与紧邻下游的第二个连续阅读框之间低效的翻译移码来表达。构建了一个融合框突变体,该突变体在移码点插入了一个额外的碱基对。这个突变体表现出高转座活性,并且应该组成性地表达转座酶InsAB'而不发生移码。出乎意料的是,从这个突变体中观察到还表达了第二种蛋白质。我们在此证明,这种蛋白质InsA*是由在修饰的移码基序上持续的移码产生的。该蛋白质保留了阻遏物InsA的活性。通过对移码基序的进一步修饰将其消除,会导致IS1的各种转座活性进一步增加。这些结果支持了这样一种假说,即单个IS1编码的蛋白质InsAB'是转座所必需的。