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Methylation-dependent transcriptional regulation of crescentin gene (creS) by GcrA in Caulobacter crescentus.
Molecular Microbiology ( IF 2.6 ) Pub Date : 2020-03-18 , DOI: 10.1111/mmi.14500 Saswat S Mohapatra 1 , Antonella Fioravanti 1 , Pauline Vandame 1 , Corentin Spriet 1 , Francesco Pini 1 , Coralie Bompard 1 , Ralf Blossey 1 , Odile Valette 2 , Emanuele G Biondi 2
Molecular Microbiology ( IF 2.6 ) Pub Date : 2020-03-18 , DOI: 10.1111/mmi.14500 Saswat S Mohapatra 1 , Antonella Fioravanti 1 , Pauline Vandame 1 , Corentin Spriet 1 , Francesco Pini 1 , Coralie Bompard 1 , Ralf Blossey 1 , Odile Valette 2 , Emanuele G Biondi 2
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In Caulobacter crescentus the combined action of chromosome replication and the expression of DNA methyl‐transferase CcrM at the end of S‐phase maintains a cyclic alternation between a full‐ to hemi‐methylated chromosome. This transition of the chromosomal methylation pattern affects the DNA‐binding properties of the transcription factor GcrA that controls the several key cell cycle functions. However, the molecular mechanism by which GcrA and methylation are linked to transcription is not fully elucidated yet. Using a combination of cell biology, genetics, and in vitro analysis, we deciphered how GcrA integrates the methylation pattern of several S‐phase expressed genes to their transcriptional output. We demonstrated in vitro that transcription of ctrA from the P1 promoter in its hemi‐methylated state is activated by GcrA, while in its fully methylated state GcrA had no effect. Further, GcrA and methylation together influence a peculiar distribution of creS transcripts, encoding for crescentin, the protein responsible for the characteristic shape of Caulobacter cells. This gene is duplicated at the onset of chromosome replication and the two hemi‐methylated copies are spatially segregated. Our results indicated that GcrA transcribed only the copy where coding strand is methylated. In vitro transcription assay further substantiated this finding. As several of the cell cycle‐regulated genes are also under the influence of methylation and GcrA‐dependent transcriptional regulation, this could be a mechanism responsible for maintaining the gene transcription dosage during the S‐phase.
中文翻译：
新月形棒状杆菌中的GcrA对新月体基因（creS）的甲基化依赖性转录调控。
在新月形杆菌中，染色体复制与S期末期DNA甲基转移酶CcrM的表达共同作用，使全甲基化至半甲基化染色体之间保持循环交替。染色体甲基化模式的这种转变会影响控制多个关键细胞周期功能的转录因子GcrA的DNA结合特性。然而，还没有完全阐明GcrA和甲基化与转录连接的分子机制。通过结合细胞生物学，遗传学和体外分析，我们解释了GcrA如何将几个S期表达基因的甲基化模式整合到它们的转录输出中。我们在体外证明了ctrA的转录Pcr启动子在半甲基化状态下被GcrA激活，而在全甲基化状态下GcrA没有作用。此外，GCRA和甲基化一起影响的特有分布CRES转录，编码crescentin，负责的特征形状的蛋白柄杆菌细胞。该基因在染色体复制开始时即被复制，并且两个半甲基化的拷贝在空间上是分开的。我们的结果表明，GcrA仅转录编码链被甲基化的拷贝。体外转录测定进一步证实了这一发现。由于一些细胞周期调控的基因也受到甲基化和GcrA依赖性转录调控的影响，这可能是在S期维持基因转录剂量的机制。
更新日期：2020-03-18
中文翻译：
新月形棒状杆菌中的GcrA对新月体基因（creS）的甲基化依赖性转录调控。
在新月形杆菌中，染色体复制与S期末期DNA甲基转移酶CcrM的表达共同作用，使全甲基化至半甲基化染色体之间保持循环交替。染色体甲基化模式的这种转变会影响控制多个关键细胞周期功能的转录因子GcrA的DNA结合特性。然而，还没有完全阐明GcrA和甲基化与转录连接的分子机制。通过结合细胞生物学，遗传学和体外分析，我们解释了GcrA如何将几个S期表达基因的甲基化模式整合到它们的转录输出中。我们在体外证明了ctrA的转录Pcr启动子在半甲基化状态下被GcrA激活，而在全甲基化状态下GcrA没有作用。此外，GCRA和甲基化一起影响的特有分布CRES转录，编码crescentin，负责的特征形状的蛋白柄杆菌细胞。该基因在染色体复制开始时即被复制，并且两个半甲基化的拷贝在空间上是分开的。我们的结果表明，GcrA仅转录编码链被甲基化的拷贝。体外转录测定进一步证实了这一发现。由于一些细胞周期调控的基因也受到甲基化和GcrA依赖性转录调控的影响，这可能是在S期维持基因转录剂量的机制。
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