Blinkowa A L, Walker J R
Department of Microbiology, University of Texas, Austin 78712.
Nucleic Acids Res. 1990 Apr 11;18(7):1725-9. doi: 10.1093/nar/18.7.1725.
The Escherichia coli dnaX gene encodes both the tau and gamma subunits of DNA polymerase III holoenzyme in one reading frame. The 71.1 kDa tau and the shorter gamma share N-terminal sequences. Mutagenesis of a potential ribosomal frameshift signal located at codons 428-430 without changing the amino acid sequence of the tau product, eliminated detectable synthesis of the gamma subunit, suggesting that the reading frame is shifted at that sequence and gamma is terminated by a nonsense codon located in the -1 frame 3 nucleotides downstream of the signal. This seems to be the first known case of a frameshift which is used, along with the termination codon in the -1 frame, to terminate a peptide within a reading frame. [Mutagenesis of a dibasic peptide (lys-lys) at codons 498-499, the site at which a tau'-'LacZ fusion protein was cleaved in vitro (1) had no effect on gamma formation in vivo, suggesting that cleavage observed in vitro is not the mechanism of gamma formation in vivo.
大肠杆菌的dnaX基因在一个阅读框中编码DNA聚合酶III全酶的tau和gamma亚基。71.1 kDa的tau与较短的gamma共享N端序列。对位于密码子428 - 430处的潜在核糖体移码信号进行诱变,且不改变tau产物的氨基酸序列,消除了可检测到的gamma亚基合成,这表明阅读框在该序列处发生移码,且gamma由位于该信号下游-1框中3个核苷酸处的一个无义密码子终止。这似乎是已知的首例移码情况,它与-1框中的终止密码子一起用于在一个阅读框内终止一个肽段。[对密码子498 - 499处的一个双碱性肽(赖氨酸-赖氨酸)进行诱变,该位点是tau'-'LacZ融合蛋白在体外被切割的位点(1),对体内gamma的形成没有影响,这表明体外观察到的切割不是体内gamma形成的机制。