Siede W, Friedl A A, Dianova I, Eckardt-Schupp F, Friedberg E C
Department of Pathology, University of Texas Southwestern Medical Center, Dallas 75235, USA.
Genetics. 1996 Jan;142(1):91-102. doi: 10.1093/genetics/142.1.91.
In mammalian cells, all subunits of the DNA-dependent protein kinase (DNA-PK) have been implicated in the repair of DNA double-strand breaks and in V(D)J recombination. In the yeast Saccharomyces cerevisiae, we have examined the phenotype conferred by a deletion of HDF1, the putative homologue of the 70-kD subunit of the DNA-end binding Ku complex of DNA-PK. The yeast gene does not play a role in radiation-induced cell cycle checkpoint arrest in G1 and G2 or in hydroxyurea-induced checkpoint arrest in S. In cells competent for homologous recombination, we could not detect any sensitivity to ionizing radiation or to methyl methanesulfonate (MMS) conferred by a hdf1 deletion and indeed, the repair of DNA double-strand breaks was not impaired. However, if homologous recombination was disabled (rad52 mutant background), inactivation of HDF1 results in additional sensitization toward ionizing radiation and MMS. These results give further support to the notion that, in contrast to higher eukaryotic cells, homologous recombination is the favored pathway of double-strand break repair in yeast whereas other competing mechanisms such as the suggested pathway of DNA-PK-dependent direct break rejoining are only of minor importance.
在哺乳动物细胞中,DNA依赖性蛋白激酶(DNA-PK)的所有亚基都与DNA双链断裂的修复以及V(D)J重组有关。在酿酒酵母中,我们研究了HDF1缺失所赋予的表型,HDF1是DNA-PK的DNA末端结合Ku复合物70-kD亚基的假定同源物。酵母基因在辐射诱导的G1和G2期细胞周期检查点停滞或羟基脲诱导的S期检查点停滞中不起作用。在有能力进行同源重组的细胞中,我们未检测到hdf1缺失导致的对电离辐射或甲基磺酸甲酯(MMS)的任何敏感性,实际上,DNA双链断裂的修复并未受损。然而,如果同源重组被禁用(rad52突变背景),HDF1的失活会导致对电离辐射和MMS的额外敏感性。这些结果进一步支持了这样一种观点,即与高等真核细胞不同,同源重组是酵母中双链断裂修复的首选途径,而其他竞争机制,如推测的DNA-PK依赖性直接断裂重新连接途径,仅具有次要重要性。