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The Heparin-binding hemagglutinin protein of Mycobacterium tuberculosis is a nucleoid-associated protein

ABSTRACT: The Heparin-binding hemagglutinin protein of Mycobacterium tuberculosis is a nucleoid-associated protein

PROVIDER: PRJNA985662 | ENA |

REPOSITORIES: ENA

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Virulence regulator EspR of Mycobacterium tuberculosis is a nucleoid-associated protein

Project description:The principal virulence determinant of Mycobacterium tuberculosis (Mtb), the ESX-1 protein secretion system, is positively controlled at the transcriptional level by EspR. Depletion of EspR reportedly affects a small number of genes, both positively or negatively, including a key ESX-1 component, the espACD operon. EspR is also thought to be an ESX-1 substrate. Using EspR-specific antibodies in ChIP-Seq experiments (chromatin immunoprecipitation followed by ultra-high throughput DNA sequencing) we show that EspR binds to at least 165 loci on the Mtb genome. Included in the EspR regulon are genes encoding not only EspA, but also EspR itself, the ESX-2 and ESX-5 systems, a host of diverse cell wall functions, such as production of the complex lipid PDIM (phenolthiocerol dimycocerosate) and the PE/PPE cell-surface proteins. EspR binding sites are not restricted to promoter regions and can be clustered. This suggests that rather than functioning as a classical regulatory protein EspR acts globally as a nucleoid-associated protein capable of long-range interactions consistent with a recently established structural model. EspR expression was shown to be growth phase-dependent, peaking in the stationary phase. Overexpression in Mtb strain H37Rv revealed that EspR influences target gene expression both positively or negatively leading to growth arrest. At no stage was EspR secreted into the culture filtrate. Thus, rather than serving as a specific activator of a virulence locus, EspR is a novel nucleoid-associated protein, with both architectural and regulatory roles, that impacts cell wall functions and pathogenesis through multiple genes. ChIP-Seq of EspR in Mtb H37Rv at mid-log phase of growth. Two independent experiments were performed. Input DNA (No IP) was used as a control.

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The Heparin-binding hemagglutinin protein of Mycobacterium tuberculosis is a nucleoid-associated protein

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Essential Nucleoid Associated Protein mIHF (Rv1388) controls virulence and housekeeping genes in Mycobacterium tuberculosis

Project description:Essential Nucleoid Associated Protein mIHF (Rv1388) controls virulence and housekeeping genes in Mycobacterium tuberculosis

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Virulence regulator EspR of Mycobacterium tuberculosis is a nucleoid-associated protein

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Chromosomal DNA binding of mIHF

Project description:mIHF is a possible nucleoid associated protein, but its DNA binding characteristics are unknown. To address this, we performed a ChIP-seq analysis of mIHF in Mycobacterium tuberculosis.

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Impact of mIHF on gene expression

Project description:mIHF is a possible nucleoid associated protein. Its effect on gene regulation remains unknown. We performed an RNA-seq analysis of the mihF conditional knock-down mutant in Mycobacterium tuberculosis. We show that mihF is essential for in vitro growth and that depeltion of mIHF results in deregulation of a large number of genes. Notably, the most downregulated gene was the espACD operon, which is necessary for secretion of M. tuberculosis main virulence factor EsxA.

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Systems-wide analysis of the GATC-binding nucleoid-associated protein Gbn and its impact on Streptomyces development

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Transcriptional profiling of Mycobacterium tuberculosis Rv3852 (hns) knockout mutant

Project description:A handful of nucleoid-associated proteins (NAPs) regulate the vast majority of genes in a bacterial cell. H-NS, the Histone-like Nucleoid-Structuring protein, is one of these NAPs and protects Escherichia coli from foreign gene expression. Though lacking any sequence similarity with E. coli H-NS, Rv3852 was annotated as the H-NS ortholog in Mycobacterium tuberculosis, as it resembles human histone H1. The role of H-NS was thoroughly investigated by immunoblotting, subcellular localization, construction of an unmarked hns deletion in the M. tuberculosis genome and subsequent analysis of the resulting Δhns strain. We found that H-NS was predominantly present in the logarithmic growth phase with a decrease in protein abundance in stationary phase. Furthermore, it was strongly associated with the cell membrane and not detected in the cytosolic fraction, nor was it secreted. The Δhns strain displayed no growth defect or morphological abnormalities. Quantitative measurement of nucleoid localization in Δhns compared to the parental H37Rv strain showed no difference in nucleoid position or spread. Infection of macrophages as well as severe combined immunodeficient (SCID) mice demonstrated that loss of H-NS had no detectable influence on the virulence of M. tuberculosis. Only few genes wre differentially expressed in the Δhns strain. We thus conclude that M. tuberculosis H-NS is not involved in pathogenesis and is not a typical NAP.

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Project description:Virulence regulator EspR of Mycobacterium tuberculosis is a nucleoid-associated protein

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