Ke N, Voytas D F
Department of Zoology and Genetics, Iowa State University, Ames 50011, USA.
Genetics. 1997 Oct;147(2):545-56. doi: 10.1093/genetics/147.2.545.
Retroelement cDNA can integrate into the genome using the element-encoded integrase or it can recombine with preexisting elements using the recombination system of the host. Recombination is a particularly important pathway for the yeast retrotransposon Ty5 and accounts for approximately 30% of the putative transposition events when a homologous substrate is carried on a plasmid and approximately 7% when the substrate is located at the chromosomal URA3 locus. Characterization of recombinants revealed that they are either simple replacements of the marker gene tandem elements. Using an assay system in which the donor element and recombination substrates are separated, we found that the long terminal repeats (LTRs) are critical for tandem element formation. LTR-containing substrates generate tandem elements at frequencies more than 10-fold higher than similarly sized internal Ty5 sequences. Internal sequences, however, facilitate tandem element formation when associated with an LTR, and there is a linear relationship between frequencies of tandem element formation and the length of LTR-containing substrates. We propose that recombination is initiated between the LTRs of the cDNA and substrate and that internal sequences promote tandem element formation by facilitating sequence alignment. Because of its location in subtelomeric regions, recombinational amplification of Ty5 may contribute to the organizations of chromosome ends.
逆转录元件cDNA可利用元件编码的整合酶整合到基因组中,或者利用宿主的重组系统与已有的元件发生重组。对于酵母逆转座子Ty5而言,重组是一条尤为重要的途径,当同源底物携带在质粒上时,约30%的推定转座事件是由重组引起的;而当底物位于染色体URA3位点时,这一比例约为7%。对重组体的表征显示,它们要么是标记基因串联元件的简单替换。通过使用一种供体元件与重组底物分离的检测系统,我们发现长末端重复序列(LTR)对于串联元件的形成至关重要。含LTR的底物产生串联元件的频率比同样大小的Ty5内部序列高出10倍以上。然而,内部序列与LTR相关联时会促进串联元件的形成,并且串联元件形成频率与含LTR底物的长度之间存在线性关系。我们提出,重组起始于cDNA和底物的LTR之间,并且内部序列通过促进序列比对来促进串联元件的形成。由于Ty5位于亚端粒区域,其重组扩增可能有助于染色体末端的组织。