Wood E R, Ghané F, Grogan D W
Department of Biological Sciences, University of Cincinnati, Ohio 54221-0006, USA.
J Bacteriol. 1997 Sep;179(18):5693-8. doi: 10.1128/jb.179.18.5693-5698.1997.
The archaea which populate geothermal environments are adapted to conditions that should greatly destabilize the primary structure of DNA, yet the basic biological aspects of DNA damage and repair remain unexplored for this group of prokaryotes. We used auxotrophic mutants of the extremely thermoacidophilic archaeon Sulfolobus acidocaldarius to assess genetic and physiological effects of a well-characterized DNA-damaging agent, short-wavelength UV light. Simple genetic assays enabled quantitative dose-response relationships to be determined and correlated for survival, phenotypic reversion, and the formation of genetic recombinants. Dose-response relationships were also determined for survival and phenotypic reversion of the corresponding Escherichia coli auxotrophs with the same equipment and procedures. The results showed S. acidocaldarius to be about twice as UV sensitive as E. coli and to be equally UV mutable on a surviving-cell basis. Furthermore, UV irradiation significantly increased the frequency of recombinants recovered from genetic-exchange assays of S. acidocaldarius. The observed UV effects were due to the short-wavelength (i.e., UV-C) portion of the spectrum and were effectively reversed by subsequent illumination of S. acidocaldarius cells with visible light (photoreactivation). Thus, the observed responses are probably initiated by the formation of pyrimidine dimers in the S. acidocaldarius chromosome. To our knowledge, these results provide the first evidence of error-prone DNA repair and genetic recombination induced by DNA damage in an archaeon from geothermal habitats.
栖息于地热环境中的古生菌适应了理应会极大破坏DNA一级结构的条件,但对于这组原核生物而言,DNA损伤与修复的基本生物学问题仍未得到探索。我们利用极端嗜热嗜酸古生菌嗜酸热硫化叶菌的营养缺陷型突变体,来评估一种特征明确的DNA损伤剂——短波长紫外光的遗传和生理效应。简单的遗传学分析能够确定定量的剂量反应关系,并将其与存活、表型回复以及基因重组体的形成相关联。利用相同的设备和程序,也确定了相应大肠杆菌营养缺陷型的存活和表型回复的剂量反应关系。结果表明,嗜酸热硫化叶菌对紫外线的敏感性约为大肠杆菌的两倍,且在存活细胞基础上对紫外线的诱变率相同。此外,紫外线照射显著增加了从嗜酸热硫化叶菌的基因交换试验中回收的重组体频率。观察到的紫外线效应归因于光谱的短波长(即UV-C)部分,并且通过随后用可见光照射嗜酸热硫化叶菌细胞(光复活)可有效逆转这种效应。因此,观察到的反应可能是由嗜酸热硫化叶菌染色体中嘧啶二聚体的形成引发的。据我们所知,这些结果首次证明了来自地热栖息地的古生菌中由DNA损伤诱导的易错DNA修复和基因重组。