Weinstock K G, Mastrangelo M F, Burkett T J, Garfinkel D J, Strathern J N
Laboratory of Eukaryotic Gene Expression, National Cancer Institute-Frederick Cancer Research Facility, Maryland 21701.
Mol Cell Biol. 1990 Jun;10(6):2882-92. doi: 10.1128/mcb.10.6.2882-2892.1990.
We have identified a novel integrated form of the yeast retrotransposon Ty consisting of multiple elements joined into large arrays. These arrays were first identified among Ty-induced alpha-pheromone-resistant mutants of MATa cells of Saccharomyces cerevisiae which contain Ty insertions at HML alpha that result in the expression of that normally silent cassette. These insertions are multimeric arrays of both the induced genetically marked Ty element and unmarked Ty elements. Structural analysis of the mutations indicated that the arrays include tandem direct repeats of Ty elements separated by only a single long terminal repeat. The Ty-HML junction fragments of one mutant were cloned and shown to contain a 5-base-pair duplication of the target sequence that is characteristic of a Ty transpositional insertion. In addition, the arrays include rearranged Ty elements that do not have normal long terminal repeat junctions. We have also identified multimeric Ty insertions at other chromosomal sites and as insertions that allow expression of a promoterless his3 gene on a plasmid. The results suggest that Ty transposition includes an intermediate that can undergo recombination to produce multimers.
我们已经鉴定出一种新型的酵母逆转录转座子Ty的整合形式,它由多个元件连接成大的阵列组成。这些阵列最初是在酿酒酵母MATa细胞的Ty诱导的α-信息素抗性突变体中发现的,这些突变体在HMLα处含有Ty插入,导致该通常沉默的基因座表达。这些插入是诱导的基因标记的Ty元件和未标记的Ty元件的多聚体阵列。对这些突变的结构分析表明,这些阵列包含仅由单个长末端重复序列分隔的Ty元件的串联直接重复序列。一个突变体的Ty-HML连接片段被克隆,并显示含有靶序列的5个碱基对重复,这是Ty转座插入的特征。此外,这些阵列包括没有正常长末端重复连接的重排Ty元件。我们还在其他染色体位点鉴定出多聚体Ty插入,以及作为允许质粒上无启动子的his3基因表达的插入。结果表明,Ty转座包括一个可以进行重组以产生多聚体的中间体。