Involvement of Transcription Elongation Factor GreA in Mycobacterium Viability, Antibiotic Susceptibility, and Intracellular Fitness

Feng, Siyuan; Liu, Yan; Liang, Wanfei; Ahmed, Mohamed Abd El-Gawad El-Sayed; Zhao, Zihan; Cong, Shan; Roberts, Adam P.; Liang, Lujie; Liao, Liya; Zhong, Zhijuan; Guo, Zhaowang; Yang, Yongqiang; Xing, Wen; Chen, Hongtao; Tian, Guo‐Bao

doi:10.3389/fmicb.2020.00413

Cited by 13 publications

(13 citation statements)

References 54 publications

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“…RNA preparation and RNA-seq analysis were performed as previously described ( Feng et al, 2020 ). Briefly, wild-type M. bovis BCG as well as the TCC deletion strain were grown to the log-phase (OD 600 , 0.5) in Middlebrook 7H9-OADC.…”

Section: Methodsmentioning

confidence: 99%

The Involvement of Mycobacterium Type III-A CRISPR-Cas System in Oxidative Stress

Yang

Liang

et al. 2021

Front. Microbiol.

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Type I and type II CRISPR-Cas systems are employed to evade host immunity by targeting interference of bacteria’s own genes. Although Mycobacterium tuberculosis (M. tuberculosis), the causative agent of tuberculosis, possesses integrated type III-A CRISPR-Cas system, its role in mycobacteria remains obscure. Here, we observed that seven cas genes (csm2∼5, cas10, cas6) were upregulated in Mycobacterium bovis BCG under oxidative stress treatment, indicating the role of type III-A CRISPR-Cas system in oxidative stress. To explore the functional role of type III-A CRISPR-Cas system, TCC (Type III-A CRISPR-Cas system, including cas6, cas10, and csm2-6) mutant was generated. Deletion of TCC results in increased sensitivity in response to hydrogen peroxide and reduced cell envelope integrity. Analysis of RNA-seq dataset revealed that TCC impacted on the oxidation-reduction process and the composition of cell wall which is essential for mycobacterial envelop integrity. Moreover, disrupting TCC led to poor intracellular survival in vivo and in vitro. Finally, we showed for the first time that TCC contributed to the regulation of regulatory T cell population, supporting a role of TCC in modulating host immunity. Our finding reveals the important role of TCC in cell envelop homeostasis. Our work also highlights type III-A CRISPR-Cas system as an important factor for intracellular survival and host immunoregulation in mycobacteria, thus may be a potential target for therapy.

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Section: Methodsmentioning

confidence: 99%

The Involvement of Mycobacterium Type III-A CRISPR-Cas System in Oxidative Stress

Yang

Liang

et al. 2021

Front. Microbiol.

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“…subtilis 168 (Revilla‐Guarinos et al, 2020); B . cereus F837/76 (Jessberger et al, 2019); Clostridium perfringens 13 (Soncini et al, 2020); Streptomyces coelicolor A3(2) (Jeong et al, 2016); Mycobacterium smegmatis MC2‐155 (Feng et al, 2020); Myxococcus xanthus DK1622 (SRA accession code: PRJNA516475); Bacteroides vulgatus ATCC8482 (SRA accession code: PRJNA473003). Reads were mapped to the respective reference genome sequences, and gene expression levels were calculated in Genious Prime 2020.2.3 (https://www.geneious.com).…”

Section: Methodsmentioning

confidence: 99%

“…Expression levels were generally highest during mid-log and transition, and lowest during late and stationary phases, with modest changes between the ratios of expression of the different gene copies at all stages. The RNA-seq data set for M. smegmatis comparing changes in gene expression on the deletion of the transcript cleavage factor GreA that is important in rescuing back-tracked RNAP (Feng et al, 2020) showed that expression of the single helD gene was substantially higher than in most other organisms, at about 25% the level of rpoB suggesting HelD may be particularly abundant in the Mycobacteria (Figure 7e). The expression levels of helD were similar in the presence and absence of greA indicating each factor acts on stalled transcription complexes independently of each other.…”

Section: S Coelicolor A2mentioning

confidence: 99%

Multiple classes and isoforms of the RNA polymerase recycling motor protein HelD

et al. 2021

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Transcription elongation is punctuated by pauses that serve important functions in permitting correct folding of structural RNA, efficient coupling of transcription and translation, and ensuring efficient transcription termination at the correct site (Saba et al., 2019). Whilst most pausing events serve an important function, on occasion RNA polymerase (RNAP) is unable to restart transcription and must be removed from the DNA to prevent damaging collisions with the DNA replication machinery or other transcription

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“…e transcript cleavage factor, GreA, interacts with the RNAP secondary channel and stimulates the intrinsic transcript cleavage activity of RNAP for the removal of the aberrant RNA 3′ ends. erefore, polymerization activity can be restarted from the end of a cleaved RNA allowing transcription to resume [43]. GreA is essential for the survival of bacteria under stress [44] and it can facilitate RecBCDmediated resection and inhibits RecA, which plays an important role in impeding DNA break repair in E. coli [45], indicating the role of GreA in adapting to the stressful environments.…”

Section: Translationmentioning

confidence: 99%

TMT-Based Quantitative Proteomics Analysis of the Fish-Borne Spoiler Shewanella putrefaciens Subjected to Cold Stress Using LC-MS/MS

Gao

Mei

et al. 2021

Journal of Chemistry

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Shewanella putrefaciens is a specific spoilage bacterium for fish during cold storage. To better understand the molecular mechanisms of cold stress adaptation of S. putrefaciens, tandem mass tag- (TMT-) based quantitative proteomic analysis was performed to detect the effects of cold stress on protein expression profiles in S. putrefaciens which had been cultivated at 4°C and 30°C, respectively. A total of 266670 peptide spectrum matching numbers were quantified proteins after data analysis. Of the 2292 proteins quantitatively analyzed, a total of 274 were found to be differentially expressed (DE) under cold stress compared with the nonstress control. By integrating the results of Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses, 9 common KEGG terms were found notable for the cold-responsive proteins. Generally, the DE proteins involved in carbohydrate, amino acid, and fatty acid biosynthesis and metabolism were significantly upregulated, leading to a specific energy conservation survival mode. The DE proteins related to DNA repair, transcription, and translation were upregulated, implicating change of gene expression and more protein biosynthesis needed in response to cold stress.

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Involvement of Transcription Elongation Factor GreA in Mycobacterium Viability, Antibiotic Susceptibility, and Intracellular Fitness

Cited by 13 publications

References 54 publications

The Involvement of Mycobacterium Type III-A CRISPR-Cas System in Oxidative Stress

The Involvement of Mycobacterium Type III-A CRISPR-Cas System in Oxidative Stress

Multiple classes and isoforms of the RNA polymerase recycling motor protein HelD

TMT-Based Quantitative Proteomics Analysis of the Fish-Borne Spoiler Shewanella putrefaciens Subjected to Cold Stress Using LC-MS/MS

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