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UV Photochemistry of DNA In Vitro and in Bacillus subtilis Spores at Earth-Ambient and Low Atmospheric Pressure: Implications for Spore Survival on Other Planets or Moons in the Solar System

机译:在地球环境和低气压下体外和枯草芽孢杆菌孢子中DNA的紫外线光化学:对太阳系中其他行星或月球上孢子存活的影响

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Two major parameters influencing the survival of Bacillus subtilis spores in space and on bodies within the Solar System are UV radiation and vacuum, both of which induce inactivating damage to DNA. To date, however, spore survival and DNA photochemistry have been explored only at the extremes of Earth-normal atmospheric pressure (101.3 kPa) and at simulated space vacuum (10~(-3)-10~(-6) Pa). In this study, wild-type spores, mutant spores lacking α/β-type small, acid-soluble spore proteins (SASP), naked DNA, and complexes between SASP SspC and DNA were exposed simultaneously to UV (254 nm) at intermediate pressure (1-2 Pa), and the UV photoproducts cis, syn-thymine-thymine cyclobutane dimer (c,sTT), trans,syn-thymine-thymine cyclobutane dimer (t,sTT), and "spore photoproduct" (SP) were quantified. At 101.3 kPa, UV-treated wild-type spores accumulated only SP, but spores treated with UV radiation at 1-2 Pa exhibited a spectrum of DNA damage similar to that of spores treated at 10~(-6) Pa, with accumulation of SP, c,sTT, and t,sTT. The presence or absence of α/β-type SASP in spores was partly responsible for the shift observed between levels of SP and c,sTT, but not t,sTT. The changes observed in spore DNA photochemistry at 1-2 Pa in vivo were not reproduced by irradiation of naked DNA or SspC:DNA complexes in vitro, suggesting that factors other than SASP are involved in spore DNA photochemistry at low pressure.
机译:影响太空中和太阳系内物体上枯草芽孢杆菌孢子存活的两个主要参数是紫外线辐射和真空,这两个参数都会导致DNA失活。然而,迄今为止,仅在地球正常大气压力(101.3 kPa)和模拟空间真空(10〜(-3)-10〜(-6)Pa)的极端条件下研究了孢子存活和DNA光化学。在这项研究中,野生型孢子,缺少α/β型小的酸溶性孢子蛋白(SASP)的突变型孢子,裸露的DNA以及SASP SspC和DNA之间的复合物在中等压力下同时暴露于紫外线(254 nm) (1-2 Pa),而UV光产物为顺式,胸腺嘧啶-胸腺嘧啶环丁烷二聚体(c,sTT),反式,胸腺嘧啶-胸腺嘧啶环丁烷二聚体(t,sTT)和“孢子光产物”(SP)为量化。在101.3 kPa下,经紫外线处理的野生型孢子仅积累SP,但是在1-2 Pa的UV辐射下处理的孢子表现出的DNA损伤谱与在10〜(-6)Pa下处理的孢子相似,但积累了SP,c,sTT和t,sTT。孢子中是否存在α/β型SASP是造成SP和c,sTT水平变化的原因,而不是t,sTT水平变化的部分原因。裸露的DNA或SspC:DNA复合物在体外照射不能重现体内1-2 Pa的孢子DNA光化学变化,这表明在低压下孢子DNA光化学中不涉及SASP。

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