Frampton Jonathan, Irmisch Anja, Green Catherine M, Neiss Andrea, Trickey Michelle, Ulrich Helle D, Furuya Kanji, Watts Felicity Z, Carr Antony M, Lehmann Alan R
Genome Damage and Stability Centre, University of Sussex, Falmer, Brighton BN1 9RQ, United Kingdom.
Mol Biol Cell. 2006 Jul;17(7):2976-85. doi: 10.1091/mbc.e05-11-1008. Epub 2006 Apr 26.
Ubiquitination of proliferating cell nuclear antigen (PCNA) plays a crucial role in regulating replication past DNA damage in eukaryotes, but the detailed mechanisms appear to vary in different organisms. We have examined the modification of PCNA in Schizosaccharomyces pombe. We find that, in response to UV irradiation, PCNA is mono- and poly-ubiquitinated in a manner similar to that in Saccharomyces cerevisiae. However in undamaged Schizosaccharomyces pombe cells, PCNA is ubiquitinated in S phase, whereas in S. cerevisiae it is sumoylated. Furthermore we find that, unlike in S. cerevisiae, mutants defective in ubiquitination of PCNA are also sensitive to ionizing radiation, and PCNA is ubiquitinated after exposure of cells to ionizing radiation, in a manner similar to the response to UV-irradiation. We show that PCNA modification and cell cycle checkpoints represent two independent signals in response to DNA damage. Finally, we unexpectedly find that PCNA is ubiquitinated in response to DNA damage when cells are arrested in G2.
增殖细胞核抗原(PCNA)的泛素化在真核生物中调控绕过DNA损伤的复制过程中起着关键作用,但具体机制在不同生物体中似乎有所不同。我们研究了粟酒裂殖酵母中PCNA的修饰情况。我们发现,在紫外线照射下,PCNA以类似于酿酒酵母中的方式发生单泛素化和多泛素化。然而,在未受损的粟酒裂殖酵母细胞中,PCNA在S期被泛素化,而在酿酒酵母中它被SUMO化。此外,我们发现,与酿酒酵母不同,PCNA泛素化缺陷的突变体对电离辐射也敏感,并且细胞暴露于电离辐射后PCNA会被泛素化,其方式类似于对紫外线照射的反应。我们表明,PCNA修饰和细胞周期检查点代表了对DNA损伤的两个独立信号。最后,我们意外地发现,当细胞停滞在G2期时,PCNA会因DNA损伤而被泛素化。