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首页> 外文期刊>Cell cycle >Rad24 truncation, coupled with altered telomere structure, promotes cdc13-1 suppression in S. cerevisiae.
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Rad24 truncation, coupled with altered telomere structure, promotes cdc13-1 suppression in S. cerevisiae.

机译：Rad24截断，再加上端粒结构的改变，促进酿酒酵母中的cdc13-1抑制。

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摘要

Distinguishing telomeres from DNA double strand breaks is critical for genome stability. In S. cerevisiae, the Cdc13 single-strand telomere binding protein is critical for protecting chromosome ends. The C-rich telomere strand is lost at high temperatures in cdc13-1 strains, leading to activation of the DNA damage checkpoint and cell inviability. Through a screen performed to identify activities involved in telomere C-strand loss, we identified two new rad24 alleles. Rad24 is an alternate Rfc1 subunit, functioning to load the 9-1-1 checkpoint clamp. In each rad24 allele, a transposon inserted within the RAD24 coding region leads to expression of different carboxyl-terminal portions of Rad24, deleting or truncating the amino-terminus. We show that an intact Rad24 amino-terminus is necessary for its checkpoint function. Interestingly, the initial cdc13-1 rad24-2 strains grew at 36 degrees C, but the extent of suppression associated with rad24-2 weakened in serial backcrosses, and cdc13-1 segregants from these crosses showed a modest increase in temperature resistance. Moreover, while a RAD24 plasmid suppressed the checkpoint defect in the initial cdc13-1 rad24-2 strain, the temperature resistance was only partially suppressed. These data suggest that the TG(1-3) amplification observed in this strain contributes to the suppression phenotype. By recreating the rad24-2 allele in a strain with normal telomeres, we find that, relative to the rad24-delta allele, rad24-2 increases the frequency of obtaining cdc13-1 cells capable of growth at high temperatures. Our hypothesis is that the Rad24-2 truncation protein affects telomere structure or recombination in a manner distinct from rad24-delta.
机译：从DNA双链断裂中区分端粒对于基因组稳定性至关重要。在酿酒酵母中，Cdc13单链端粒结合蛋白对于保护染色体末端至关重要。富含C的端粒链在高温下会在cdc13-1菌株中丢失，从而导致DNA损伤检查点的激活和细胞的生存能力。通过执行筛选以鉴定涉及端粒C链丢失的活动，我们鉴定了两个新的rad24等位基因。 Rad24是另一个Rfc1子单元，用于加载9-1-1检查点钳位。在每个rad24等位基因中，插入RAD24编码区域内的转座子导致Rad24的不同羧基末端部分表达，缺失或截短氨基末端。我们表明，完整的Rad24氨基末端是其检查点功能所必需的。有趣的是，最初的cdc13-1 rad24-2菌株在36°C时生长，但是在连续回交中与rad24-2相关的抑制程度减弱了，这些交叉的cdc13-1分离子显示出适度的耐温性提高。此外，虽然RAD24质粒抑制了初始cdc13-1 rad24-2菌株中的检查点缺陷，但耐热性仅被部分抑制。这些数据表明，在该菌株中观察到的TG（1-3）扩增有助于抑制表型。通过在具有正常端粒的菌株中重建rad24-2等位基因，我们发现，相对于rad24-delta等位基因，rad24-2增加了获得能够在高温下生长的cdc13-1细胞的频率。我们的假设是Rad24-2截短蛋白以不同于rad24-delta的方式影响端粒结构或重组。

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