Schons-Fonseca Luciane, da Silva Josefa B, Milanez Juliana S, Domingos Renan H, Smith Janet L, Nakaya Helder I, Grossman Alan D, Ho Paulo L, da Costa Renata M A
Centro de Biotecnologia, Instituto Butantan, São Paulo, São Paulo 05503-900, Brazil Instituto de Química, Universidade de São Paulo, São Paulo, São Paulo 05508-000, Brazil Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts, MA 02139, USA.
Centro de Biotecnologia, Instituto Butantan, São Paulo, São Paulo 05503-900, Brazil.
Nucleic Acids Res. 2016 Feb 18;44(3):1179-91. doi: 10.1093/nar/gkv1536. Epub 2016 Jan 13.
We determined the effects of DNA damage caused by ultraviolet radiation on gene expression in Leptospira interrogans using DNA microarrays. These data were integrated with DNA binding in vivo of LexA1, a regulator of the DNA damage response, assessed by chromatin immunoprecipitation and massively parallel DNA sequencing (ChIP-seq). In response to DNA damage, Leptospira induced expression of genes involved in DNA metabolism, in mobile genetic elements and defective prophages. The DNA repair genes involved in removal of photo-damage (e.g. nucleotide excision repair uvrABC, recombinases recBCD and resolvases ruvABC) were not induced. Genes involved in various metabolic pathways were down regulated, including genes involved in cell growth, RNA metabolism and the tricarboxylic acid cycle. From ChIP-seq data, we observed 24 LexA1 binding sites located throughout chromosome 1 and one binding site in chromosome 2. Expression of many, but not all, genes near those sites was increased following DNA damage. Binding sites were found as far as 550 bp upstream from the start codon, or 1 kb into the coding sequence. Our findings indicate that there is a shift in gene expression following DNA damage that represses genes involved in cell growth and virulence, and induces genes involved in mutagenesis and recombination.
我们使用DNA微阵列确定了紫外线辐射引起的DNA损伤对问号钩端螺旋体基因表达的影响。这些数据与LexA1（一种DNA损伤反应调节因子）在体内的DNA结合情况相结合，通过染色质免疫沉淀和大规模平行DNA测序（ChIP-seq）进行评估。对DNA损伤作出反应时，钩端螺旋体诱导了参与DNA代谢、移动遗传元件和缺陷原噬菌体的基因表达。参与去除光损伤的DNA修复基因（如核苷酸切除修复uvrABC、重组酶recBCD和解离酶ruvABC）未被诱导。参与各种代谢途径的基因被下调，包括参与细胞生长、RNA代谢和三羧酸循环的基因。从ChIP-seq数据中，我们在1号染色体上观察到24个LexA1结合位点，在2号染色体上观察到1个结合位点。DNA损伤后，这些位点附近的许多（但不是全部）基因的表达增加。结合位点在起始密码子上游550 bp处或编码序列内1 kb处被发现。我们的研究结果表明，DNA损伤后基因表达发生了变化，抑制了参与细胞生长和毒力的基因，并诱导了参与诱变和重组的基因。
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