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Molecular switching of a DNA-sliding clamp to a repressor mediates long-range gene silencing

ABSTRACT: Molecular switching of a DNA-sliding clamp to a repressor mediates long-range gene silencing

PROVIDER: PRJNA1147220 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR30230767_1.fastq.gz	Fastqsanger.gz
	SRR30230767_2.fastq.gz	Fastqsanger.gz
	SRR30230768_1.fastq.gz	Fastqsanger.gz
	SRR30230768_2.fastq.gz	Fastqsanger.gz
	SRR30230769_1.fastq.gz	Fastqsanger.gz

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Molecular switching of a DNA-sliding clamp to a repressor mediates long-range gene silencing

Project description:Long-range gene regulation is rare in bacteria and is confined to the classical DNA looping model. Here, we use a combination of biophysical approaches, including X-ray crystallography and single-molecule analysis, to show that long-range gene silencing on the plasmid RK2, a source of multidrug resistance across diverse Gram-negative bacteria, is achieved cooperatively by a DNA-sliding clamp, KorB, and a clamp-locking protein, KorA. We find that KorB is a CTPase clamp that can entrap and slide along DNA to reach distal target promoters. We resolved the tripartite crystal structure of a KorB-KorA-DNA co-complex, revealing that KorA latches KorB into a closed-clamp state. KorA thus stimulates repression by stalling KorB sliding at target promoters to occlude RNA polymerase holoenzymes. Altogether, our findings explain the mechanistic basis for KorB role-switching from a DNA-sliding clamp to a co-repressor, and provide a new paradigm for the long-range regulation of gene expression.

2024-09-01 | GSE274567 | GEO

KorB switching from DNA-sliding clamp to repressor mediates long-range gene silencing in a multi-drug resistance plasmid.

Project description:Examples of long-range gene regulation in bacteria are rare and generally thought to involve DNA looping. Here, using a combination of biophysical approaches including X-ray crystallography and single-molecule analysis for the KorB-KorA system in Escherichia coli, we show that long-range gene silencing on the plasmid RK2, a source of multi-drug resistance across diverse Gram-negative bacteria, is achieved cooperatively by a DNA-sliding clamp, KorB, and a clamp-locking protein, KorA. We show that KorB is a CTPase clamp that can entrap and slide along DNA to reach distal target promoters up to 1.5 kb away. We resolved the tripartite crystal structure of a KorB-KorA-DNA co-complex, revealing that KorA latches KorB into a closed clamp state. DNA-bound KorA thus stimulates repression by stalling KorB sliding at target promoters to occlude RNA polymerase holoenzymes. Together, our findings explain the mechanistic basis for KorB role switching from a DNA-sliding clamp to a co-repressor and provide an alternative mechanism for long-range regulation of gene expression in bacteria.

| S-EPMC11790492 | biostudies-literature

The zinc finger protein CLAMP promotes long-range chromatin interactions that mediate dosage compensation of the Drosophila male X-chromosome [4C-seq]

Project description:Hi-C and 4C-seq anlaysis of Drosophila S2 cells Drosophila dosage compensation is an important model system for defining how active chromatin domains are formed. The Male-specific lethal dosage compensation complex (MSLc) increases transcript levels of genes along the length of the single male X-chromosome to equalize with that on the two female X-chromosomes. The strongest binding sites for MSLc cluster together in three-dimensional space independent of MSLc because clustering occurs in both sexes. CLAMP, a non-sex specific, ubiquitous zinc finger protein, binds synergistically with MSLc to enrich the occupancy of both factors on the male X-chromosome. Here, we demonstrate that CLAMP promotes the observed clustering of MSLc bindings sites. Genome-wide, CLAMP promotes interactions between active chromatin regions and represses interactions between inactive chromatin regions. Moreover, the X-enriched CLAMP protein promotes longer-range interactions on the active X-chromosome than autosomes. Overall, we define how long-range interactions, mediated by a locally-enriched ubiquitous transcription factor, generate a three-dimensional active chromatin domain.

2019-12-01 | GSE130544 | GEO

The zinc finger protein CLAMP promotes long-range chromatin interactions that mediate dosage compensation of the Drosophila male X-chromosome [Hi-C seq]

2019-12-01 | GSE130545 | GEO

Peptide-Based Covalent Inhibitors Bearing Mild Electrophiles to Target a Conserved His Residue of the Bacterial Sliding Clamp

Project description:Bacterial infections caused by drug-resistant pathogens represent a major threat against which effective antibiotics acting via new mechanisms and engaging new biological targets are highly needed. The sliding clamp (SC), an essential component of the bacterial DNA replication machinery, has been identified as a new target. This processivity factor operates through multiple protein-protein interactions with conserved peptide motifs in multiple proteins involved in DNA replication and repair. Herein, we report the structure-guided design of targeted covalent inhibitors (TCIs) able to bind covalently and selectively to the E. coli SC (EcSC). The inhibitors target His175, a well-conserved SC residue located on the edge of the peptide-binding pocket. Thermodynamic and kinetic analyses confirmed the higher efficiency of the chloroacetamide warhead compared to Michael acceptors. High-resolution X-ray structures of two different covalent inhibitor–SC adducts revealed the canonical ligand orientation and confirmed the formation of the covalent bond with His175. Proteomic studies were consistent with a selective EcSC engagement by the chloroacetamide-based TCI. Finally, the TCI of SC was substantially more active than the parent non-covalent inhibitor in an in vitro SC-dependent replicative assay, suggesting the potential of the approach to target the SC of priority pathogens.

2024-01-29 | PXD042384 | Pride

The CoREST Repressor Complex Mediates Phenotype Switching and Therapy Resistance in Melanoma

Project description:The CoREST Repressor Complex Mediates Phenotype Switching and Therapy Resistance in Melanoma

| PRJNA1071546 | ENA

The CoREST Repressor Complex Mediates Phenotype Switching and Therapy Resistance in Melanoma

Project description:The CoREST Repressor Complex Mediates Phenotype Switching and Therapy Resistance in Melanoma

| PRJNA657192 | ENA

DNA binding activities of the C-clamp

Project description:Investigated the DNA binding specificity of the C-clamp of dnTCF1E Used a doxycycline inducible system to investigate the role of the C-clamp in the genome-wide binding of dnTCF1E. Induced dnTCF1EWT and dnTCF1Emut (C-clamp mutant that is null for C-clamp DNA binding) over 2 time points (2 hours and 9 hours). A 0 hour (no doxycycline, uninduced) control was also included.

2014-12-04 | E-GEOD-53536 | biostudies-arrayexpress

Sliding of Bacillus subtilis Natto

Project description:Transcriptome comparison of Bacillus subtilis Natto under sliding permissive (0.7% agar) and restrictive (1.5% agar or spo0A mutant strain) conditions.

2015-08-04 | GSE43840 | GEO

CLAMP Protein Binding Microarray (PBM) experiment

Project description:To investigate the DNA binding specificity of the CLAMP protein, we have designed a custom PBM to interrogate the binding of CLAMP to DNA sequences extracted from the Drosophila melanogaster genome. Specific regions were extracted based on ChIP-seq data, motif occurence and proximity to gene transcription start sites. N-terminal GST-tagged protein samples were made for the C-terminal four and six zinc finger portions of the protein CLAMP; samples were made by in vitro transcription translation (IVT). IVT reaction mixtures for the two CLAMP constructs were applied directly to the PBM microarray and incubated for 1hour. Microarray-bound protein was fluorescently labeled using Alexa488-conjugated antibodies targeting GST, and the microarray was scanned in using a standard microarray scanner. Median fluorescence intensity over eight replicate probes was reported for each unique DNA sequence on the microarray.

2016-07-17 | E-GEOD-83442 | biostudies-arrayexpress

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