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Mutator-Like Element in the Yeast Yarrowia lipolytica Displays Multiple Alternative Splicings

机译:酵母解脂耶氏酵母中的突变体样元素显示多个替代剪接。

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摘要

A new type of DNA transposon, Mutyl, has been identified in the sequenced genome of the yeast Yarrowia lipolytica. This transposon is 7,413 bp long and carries two open reading frames (ORFs) which potentially encode proteins of 459 and 1,178 amino acids, respectively. Whereas the first ORF shows no significant homology to previously described proteins, the second ORF shows sequence similarities with various Mutator-like element (MULE)-encoded transposases, including the bacterial transposase signature sequence. Other MULE features shared by Mutyl include a zinc finger motif in the putative transposase, a 22-bp-long imperfect inverted repeat at each end, and a 9- to 10-bp duplication of its target site in the chromosome. Of the five copies of Mutyl present in the genome, one has a deletion of the first 8 bases, and the others are full length with a single base change in one element. The first potential gene of Mutyl, mutB, was shown to be expressed in exponentially growing cells. Its sequence contains a predicted intron with two 5′ splice sites, a single branch point, and two 3′ splice sites. Its mRNA is alternatively spliced, as judged by reverse transcription-PCR, and generates four mRNAs corresponding to protein-coding sequences of 128, 156, 161, and 190 amino acids. Of the three distinct lineages characterized in Y. lipolytica, strains from the German lineage and the French lineage do not carry Mutyl. A study of the distribution of Mutyl in strains of the French lineage evidenced a recent transposition event. Taken together, these results indicate that Mutyl is still active.
机译:在酵母解脂耶氏酵母的测序基因组中已鉴定出一种新型的DNA转座子,Mutyl。该转座子长7,413 bp,带有两个开放阅读框(ORF),分别可能编码459和1,178个氨基酸。尽管第一个ORF与先前描述的蛋白质没有显着的同源性,但是第二个ORF与各种Mutator样元件(MULE)编码的转座酶显示了序列相似性,包括细菌转座酶签名序列。 Mutyl共享的其他MULE功能包括推定的转座酶中的锌指基序,在每个末端的22 bp长的不完美反向重复以及其在染色体中靶位的9至10 bp重复。在基因组中存在的5个Mutyl拷贝中,一个拷贝具有前8个碱基的缺失,而其他拷贝则是全长,其中一个元素只有一个碱基的改变。 Mutyl的第一个潜在基因mutB已显示在成倍增长的细胞中表达。其序列包含具有两个5'剪接位点,一个分支点和两个3'剪接位点的预测内含子。如通过逆转录PCR所判断,其mRNA被选择性剪接,并产生对应于128、156、161和190个氨基酸的蛋白质编码序列的四个mRNA。在以解脂耶氏酵母为特征的三个不同谱系中,来自德国谱系和法国谱系的菌株不携带Mutyl。对法国谱系菌株中Mutyl分布的研究证明了最近的转座事件。综上所述,这些结果表明Mutyl仍然具有活性。

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