Eide D, Anderson P
Department of Genetics, University of Wisconsin, Madison 53706.
Mol Cell Biol. 1988 Feb;8(2):737-46. doi: 10.1128/mcb.8.2.737-746.1988.
The transposable element Tc1 is responsible for most spontaneous mutations that occur in Caenorhabditis elegans variety Bergerac. We investigated the genetic and molecular properties of Tc1 transposition and excision. We show that Tc1 insertion into the unc-54 myosin heavy-chain gene was strongly site specific. The DNA sequences of independent Tc1 insertion sites were similar to each other, and we present a consensus sequence for Tc1 insertion that describes these similarities. We show that Tc1 excision was usually imprecise. Tc1 excision was imprecise in both germ line and somatic cells. Imprecise excision generated novel unc-54 alleles that had amino acid substitutions, amino acid insertions, and, in certain cases, probably altered mRNA splicing. The DNA sequences remaining after Tc1 somatic excision were the same as those remaining after germ line excision, but the frequency of somatic excision was at least 1,000-fold higher than that of germ line excision. The genetic properties of Tc1 excision, combined with the DNA sequences of the resulting unc-54 alleles, demonstrated that excision was dependent on Tc1 transposition functions in both germ line and somatic cells. Somatic excision was not regulated in the same strain-specific manner as germ-line excision was. In a genetic background where Tc1 transposition and excision in the germ line was not detectable, Tc1 excision in the soma still occurred at high frequency.
转座因子Tc1是秀丽隐杆线虫Bergerac变种中大多数自发突变的原因。我们研究了Tc1转座和切除的遗传及分子特性。我们发现Tc1插入到unc-54肌球蛋白重链基因中具有很强的位点特异性。独立的Tc1插入位点的DNA序列彼此相似,我们给出了一个描述这些相似性的Tc1插入共有序列。我们发现Tc1切除通常是不精确的。Tc1在生殖细胞和体细胞中的切除都是不精确的。不精确切除产生了具有氨基酸替换、氨基酸插入的新的unc-54等位基因,在某些情况下,可能还改变了mRNA剪接。Tc1体细胞切除后剩余的DNA序列与生殖细胞切除后剩余的相同,但体细胞切除的频率比生殖细胞切除至少高1000倍。Tc1切除的遗传特性,结合所产生的unc-54等位基因的DNA序列,表明切除在生殖细胞和体细胞中都依赖于Tc1转座功能。体细胞切除不像生殖细胞切除那样以相同的菌株特异性方式受到调控。在一个无法检测到生殖细胞中Tc1转座和切除的遗传背景下,体细胞中的Tc1切除仍以高频发生。