Lee Julia Y, Mogen Jonathan L, Chavez Alejandro, Johnson F Brad
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
J Biol Chem. 2008 Oct 31;283(44):29847-58. doi: 10.1074/jbc.M804760200. Epub 2008 Aug 29.
In yeast telomerase mutants, the Sgs1 RecQ helicase slows the rate of senescence and also facilitates the appearance of certain types of survivors of critical telomere shortening via mechanisms dependent on Rad52-dependent homologous recombination (HR). Here we describe a third function for Sgs1 in telomerase-deficient cells, inhibition of survivors that grow independent of Rad52. Unlike tlc1 rad52 double mutants, which do not form survivors of telomere dysfunction, tlc1 rad52 sgs1 triple mutants readily generated survivors. After emerging from growth crisis, the triple mutants progressively lost telomeric and subtelomeric sequences, yet grew for more than 1 year. Analysis of cloned chromosome termini and of copy number changes of loci genome-wide using tiling arrays revealed terminal deletions extending up to 57 kb, as well as changes in Ty retrotransposon copy numbers. Amplification of the remaining terminal sequences generated large palindromes at some chromosome termini. Sgs1 helicase activity but not checkpoint function was essential for inhibiting the appearance of the survivors, and the continued absence of Sgs1 was required for the growth of the established survivors. Thus, in addition to facilitating the maintenance of telomere repeat sequences via HR-dependent mechanisms, a RecQ helicase can prevent the adoption of HR-independent mechanisms that stabilize chromosome termini without the use of natural telomere sequences. This provides a novel mechanism by which RecQ helicases may help maintain genome integrity and thus prevent age-related diseases and cancer.
在酵母端粒酶突变体中,Sgs1 RecQ解旋酶减缓衰老速率,还通过依赖于Rad52依赖的同源重组(HR)的机制促进某些类型的关键端粒缩短幸存者的出现。在此,我们描述了Sgs1在端粒酶缺陷细胞中的第三种功能,即抑制不依赖Rad52生长的幸存者。与不形成端粒功能障碍幸存者的tlc1 rad52双突变体不同,tlc1 rad52 sgs1三突变体很容易产生幸存者。从生长危机中出现后,三突变体逐渐丢失端粒和亚端粒序列,但仍生长了1年多。使用平铺阵列分析克隆的染色体末端和全基因组位点的拷贝数变化,发现末端缺失延伸至57 kb,以及Ty逆转座子拷贝数的变化。剩余末端序列的扩增在一些染色体末端产生了大的回文序列。Sgs1解旋酶活性而非检查点功能对于抑制幸存者的出现至关重要,并且已建立的幸存者的生长需要持续缺失Sgs1。因此,除了通过依赖HR的机制促进端粒重复序列的维持外,一种RecQ解旋酶可以防止采用不依赖HR的机制,这些机制在不使用天然端粒序列的情况下稳定染色体末端。这提供了一种新机制,通过该机制RecQ解旋酶可能有助于维持基因组完整性,从而预防与年龄相关的疾病和癌症。