紫外辐照恢复早期耐辐射奇球菌的转录组学分析
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华中农业大学信息学院,农业生物信息湖北省重点实验室,武汉 430070

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Q7;Q93

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国家自然科学基金(31971184)资助项目。


Transcriptomic Analysis of Deinococcus radiodurans During The Early Recovery Stage From Ultraviolet Irradiation
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Hubei Key Laboratory of Agricultural Bioinformatics, College of Informatics, Huazhong Agricultural University, Wuhan 430070, China

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This work was supported by a grant from The National Natural Science Foundation of China (31971184).

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    摘要:

    目的 耐辐射奇球菌是一种对紫外线、电离、干燥和化学试剂具有较强抗性的极端微生物。然而,该菌在紫外辐照后恢复早期的分子响应还不完全清楚。本文的目的是揭示耐辐射奇球菌在这一阶段的转录组响应。方法 本研究采用 RNA-seq技术,测定了正常和紫外辐照培养条件下耐辐射奇球菌的转录组。为确定关键的差异表达基因及其调控关系,进行了功能富集分析。选取部分关键差异表达基因,进行实时定量PCR实验验证。利用以往研究中的转录组数据,寻找紫外辐照、电离辐射和干燥胁迫条件下公共的差异表达基因。构建了蛋白质-蛋白质相互作用网络;对蛋白质互作网络中的枢纽基因和主要模块进行了鉴定;对这些枢纽基因和模块进行了功能富集分析。结果 紫外辐照后的恢复早期,上调基因数量是下调基因数量的2倍以上,且多数与应激反应和DNA修复有关。恢复早期的修复途径主要有单链退火(SSA)途径(涉及基因:ddrA-D)、非同源端连接(NHEJ)途径(涉及基因:ligBpprA)和核苷酸切除修复(NER)途径(涉及基因:uvrA-C),前两种途径为同源重组(HR)做准备,而NER途径去除紫外线照射带来的嘧啶二聚体。通过比较紫外辐照、电离辐射和干燥胁迫下的转录组数据,发现公共的差异表达基因主要涉及奇球菌属特异性基因和DNA/RNA代谢相关基因。从差异表达基因的蛋白质互作网络中发现了几个重要的枢纽基因和相互作用模块,其功能集中在双链断裂修复、DNA拓扑改变和复制。结论 在紫外辐照后的早期恢复阶段,耐辐射奇球菌的多种基因发生了转录组水平的分子响应,并启动了多个修复途径来应对这种胁迫,其中一些修复途径在其他应激条件下同样存在。

    Abstract:

    Objective Deinococcus radiodurans (D. radiodurans) is an extremophile with strong resistance to ultraviolet (UV), ionization, desiccation and chemical reagents. However, the molecular responses of this bacterium in the early recovery stage after UV irradiation are not fully understood. The aim of this work is to reveal the transcriptomic responses of D. radiodurans at this stage.Methods In this study, the transcriptomes of D. radiodurans under normal and UV irradiation culture conditions were determined by using RNA-seq technique. To identify the key genes and their regulatory relationships among the differentially expressed genes (DEGs), functional enrichment analysis was performed. Some key DEGs were selected and validated by real-time quantitative PCR. The transcriptome data from previous studies were adopted to find DEGs common to UV irradiation, ionizing radiation and desiccation stresses. The protein-protein interaction (PPI) network of DEGs was constructed; the hub genes and major modules in the PPI network were identified; functional enrichment analysis was performed for these hubs and modules.Results The results showed that the number of up-regulated genes was more than twice that of down-regulated genes in the early recovery stage after UV irradiation, and most of them were related to stress response and DNA repair. The main repair pathways in the early stage of recovery include single-strand annealing (SSA) pathway (involving genes ddrA-D), nonhomologous end joining (NHEJ) pathway (involving genes ligB and pprA) and nucleotide excision repair (NER) pathway (involving genes uvrA-C), the first two of which are for homologous recombination (HR), while the NER pathway removes pyrimidine dimers caused by UV irradiation. By comparing the transcriptome data under UV irradiation, ionizing radiation and desiccation stresses, it was found that the common responsive DEGs mainly involve Deinococcus-specific genes and the genes related to DNA/RNA metabolism. Several important hub genes and interaction modules were identified from the PPI network of DEGs, whose functions are concentrated in double-strand break repair, DNA topological change and replication.Conclusion These results indicate that in the early recovery stage after UV irradiation, a variety of genes in D. radiodurans undergo responses at transcriptome level, several repair pathways are initiated to cope with this stress, and some repair pathways are common to other stress conditions.

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张彩云,邱秦天,马彬广.紫外辐照恢复早期耐辐射奇球菌的转录组学分析[J].生物化学与生物物理进展,2023,50(7):1701-1715

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历史
  • 收稿日期:2022-10-01
  • 最后修改日期:2023-06-04
  • 接受日期:2022-12-01
  • 在线发布日期: 2023-07-19
  • 出版日期: 2023-07-20