A specific terminal structure is required for Ty1 transposition.
A specific terminal structure is required for Ty1 transposition.
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Ty1 转座需要特定的末端结构。
DOI：
10.1101/gad.4.3.324
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发表时间：
1990
影响因子：
10.5
通讯作者：
Boeke,JD
中科院分区：
文献类型：
--
作者：
Eichinger,DJ;Boeke,JD
酵母逆转录转座子 Ty1 指导由 Ty1 编码的蛋白质、RNA 和逆转录物组成的病毒样颗粒 (VLP) 的合成。 Ty1 逆转录物带有选择性标记并在 VLP 中发现，能够在体外转座到裸露的靶 DNA 中。构建了由Ty长末端重复序列(LTR)或δ组成的盒，其用supF标记，并且侧翼是适当的限制性位点。这些人工底物的末端类似于线性全长 Ty1 逆转录物的末端，可以与 VLP（包含未标记的逆转录物）共孵育，从而实现人工底物的非常有效的整合。结果表明 Ty DNA 限制了体内转座，表明低效的逆转录调节 Ty1 转座。对这些模型底物的转座进行分析表明，Ty 转座是通过逆转录物的 3' 羟基残基与靶 DNA 的连接来进行的，这些底物类似于体内 Ty1 转座中间体或与它们有细微的差异。
Yeast retrotransposon Ty1 directs the synthesis of virus-like particles (VLPs) consisting of Ty1-encoded proteins, RNA, and reverse transcripts. Ty1 reverse transcripts, tagged with a selectable marker and found within VLPs, are capable of transposing into naked target DNA in vitro. Cassettes consisting of a Ty long terminal repeat (LTR), or delta, marked with supF, and flanked by appropriate restriction sites were constructed. These artificial substrates, whose termini resemble those of linear, full-length Ty1 reverse transcripts, can be coincubated with VLPs (containing unmarked reverse transcripts), resulting in the very efficient integration of the artificial substrate. The results suggest that Ty DNA is limiting for transposition in vivo, suggesting that inefficient reverse transcription regulates Ty1 transposition. Analysis of the transposition of these model substrates, which resemble in vivo Ty1 transposition intermediates or differ from them in subtle ways, shows that Ty transposition proceeds by the linkage of the 3' hydroxyl residue of the reverse transcript to target DNA.