MvaT Family Proteins Encoded on IncP-7 Plasmid pCAR1 and the Host Chromosome Regulate the Host Transcriptome Cooperatively but Differently
MvaT Family Proteins Encoded on IncP-7 Plasmid pCAR1 and the Host Chromosome Regulate the Host Transcriptome Cooperatively but Differently
复制标题
IncP-7 质粒 pCAR1 和宿主染色体上编码的 MvaT 家族蛋白协同但不同地调节宿主转录组
DOI：
10.1128/aem.03071-15
复制
发表时间：
2015
影响因子：
4.4
通讯作者：
H. Nojiri
中科院分区：
文献类型：
--
作者：
C. Yun;Yurika Takahashi;M. Shintani;T. Takeda;C. Suzuki;K. Okada;H. Yamane;H. Nojiri
摘要MvaT蛋白是假单胞菌中H-NS蛋白家族的成员。IncP-7接合质粒pCAR 1携带mvaT同源基因pmr。在携带pCAR 1的恶臭假单胞菌KT 2440中，pmr和染色体携带的同源基因turA和turB以高水平转录，并且Pmr在体外与TurA和TurB相互作用。在本研究中，我们阐明了三种MvaT蛋白如何调节恶臭假单胞菌KT 2440（pCAR 1）的转录组。采用微阵列技术（ChIP-芯片）通过改良染色质免疫沉淀试验进行的分析表明，恶臭假单胞菌KT 2440（pCAR 1）基因组中Pmr、TurA和TurB的结合区域几乎相同;然而，在对数生长期和早期稳定生长期使用编码MvaT蛋白的基因缺失的突变体进行的转录组学分析清楚地表明，它们的调节子不同。事实上，Pmr和其他两种蛋白质之间发现了显着的调节子相异性。Pmr缺失在稳定期早期比对数期影响更多基因的转录，表明Pmr能更有效地改善pCAR 1对宿主适应性的影响。另外，TurA和TurB调节子的相似性意味着它们可能作为响应质粒携带的全局转录调节子发挥互补作用。
ABSTRACT MvaT proteins are members of the H-NS family of proteins in pseudomonads. The IncP-7 conjugative plasmid pCAR1 carries an mvaT-homologous gene, pmr. In Pseudomonas putida KT2440 bearing pCAR1, pmr and the chromosomally carried homologous genes, turA and turB, are transcribed at high levels, and Pmr interacts with TurA and TurB in vitro. In the present study, we clarified how the three MvaT proteins regulate the transcriptome of P. putida KT2440(pCAR1). Analyses performed by a modified chromatin immunoprecipitation assay with microarray technology (ChIP-chip) suggested that the binding regions of Pmr, TurA, and TurB in the P. putida KT2440(pCAR1) genome are almost identical; nevertheless, transcriptomic analyses using mutants with deletions of the genes encoding the MvaT proteins during the log and early stationary growth phases clearly suggested that their regulons were different. Indeed, significant regulon dissimilarity was found between Pmr and the other two proteins. Transcription of a larger number of genes was affected by Pmr deletion during early stationary phase than during log phase, suggesting that Pmr ameliorates the effects of pCAR1 on host fitness more effectively during the early stationary phase. Alternatively, the similarity of the TurA and TurB regulons implied that they might play complementary roles as global transcriptional regulators in response to plasmid carriage.