The role of SIR4 in Ty1 retrotransposition

描述(申请人提供)：酵母转座的TYL逆转录元件 通过一种与感染更大病毒的逆转录病毒有许多相似之处的机制 真核生物。TYL是研究病毒机制的一个很好的模型系统，因为 TYL转位途径的分离是由EASE和 酵母分子遗传技术的灵活性。TYL转位 过程本身就是温度敏感的。删除SIR4，一个沉默的 信息调节基因，在不允许的情况下部分恢复转座 温度。 SIR4与其他沉默信息调节基因的复合体 通过创建沉默的基因来转录调节交配基因和端粒 染色质区域。SIR4也被证明在细胞衰老中发挥作用。一个 SIR4的等位基因称为sir4-42，是作为一种抗逆突变体被分离出来的 还显示出更长的寿命潜力。 这笔赠款的具体目的是为了确定 SIR4在TYL转位中的作用实验将集中在 确定增加的换位事件是否发生在SIR4中 高温下的缺失应变是真正的整合事件，或者是由于 TYL基因与基因组重组的增加。Sir4基因对大鼠血管紧张素转换酶的影响 靶向将通过克隆和测序转座事件来评估。 平行实验将集中在确定Sir4p是否与TYL共净化 病毒样颗粒(VLP)以及蛋白质的加工、cDNA的生产 以及TYL在sir4突变株中的整合步骤。SIR4已广泛应用于 以及基因产物突变对TYL转座的影响 已知与sir4p相互作用或受sir4突变影响的人也会 下定决心。一种简单的基因测试将被用来确定是否高 温度会影响静音。

DESCRIPTION (provided by applicant): The Tyl retroelement of yeast transposes via a mechanism bearing many similarities to retroviruses that infect larger eukaryotes. Tyl is a good model system for studying viral mechanisms, as dissection of the Tyl transposition pathway is facilitated by the ease and flexibility of yeast molecular genetic techniques. The Tyl transposition process is inherently temperature sensitive. Deletion of SIR4, a silent information regulator gene, partially restores transposition at non-permissive temperature. SIR4 complexes with other Silent Information Regulator genes do transcriptionally regulate the mating loci and telomeres by creating a silenced chromatin region. SIR4 has also been shown to play a role in cellular aging. An allele of SIR4, called sir4-42, was isolated as a stress-resistant mutant that also demonstrates increased life span potential. The specific aims of this grant are directed towards the goal of determining the role of SIR4 in the transposition of Tyl. Experiments will focus on determining whether the increased transposition events occurring in a sir4 deletion strain at high temperature are true integration events, or are due to an increase in recombination of Tyl cDNA to the genome. The effects of sir4 on targeting will be assessed by cloning and sequencing transposition events. Parallel experiments will focus on determining if Sir4p co-purifies with Tyl Virus-Like Particles (VLPs), as well as the protein processing, cDNA production and integration steps of Tyl in sir4 mutant strains. SIR4 has been widely studied, and the effects on Tyl transposition of mutations in gene products known to interact with Sir4p or to be affected by sir4 mutations will also be determined. A simple genetic assay will be used to determine if high temperature affects silencing.