Dataset Information

Bidirectional signaling between nucleotide and substrate binding sites in a type IV multidrug ABC transporter

ABSTRACT: ATP-binding cassette (ABC) transporters use ATP to transport substrates across cellular membranes. In type IV ABC transporters, which includes many multidrug resistance (MDR) pumps, communication between the nucleotide-binding domains (NBDs) and transmembrane domains (TMDs) occurs via intracellular coupling helices. However, the precise mechanism of interdomain crosstalk and coordination between ATP and substrate binding sites remain unclear. Here, we combined NMR spectroscopy, Hydrogen-Deuterium eXchange Mass Spectrometry (HDX-MS), PET-FCS and functional assays to identify a conserved residue cluster at the NBD/TMD interface of the bacterial MDR transporter BmrA. This cluster functions as a bidirectional relay to convey nucleotide and substrate occupancy between the two domains through coupling helix 2. Mutations impact both local and global transporter dynamics. Our findings reveal a novel interdomain communication pathway in type IV ABC transporters, offering insights into the coordination of ligand binding in this important protein family.

INSTRUMENT(S):

ORGANISM(S): Escherichia Coli

SUBMITTER: Julien Marcoux

LAB HEAD: Julien Marcoux

PROVIDER: PXD058013 | Pride | 2025-11-13

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	BmrA-His-FL-MSP-Pepsin.fasta	Fasta
	BmrA-W413F-Nanodiscs-Apo-vs-ATP-Vi-vs-Doxo-Cluster.csv	Csv
	BmrA-W413F-Nanodiscs-Apo-vs-ATP-Vi-vs-Doxo-State.csv	Csv
	BmrA-WT-Nanodiscs-Apo-vs-ATP-Vi-vs-Doxo-Cluster.csv	Csv
	BmrA-WT-Nanodiscs-Apo-vs-ATP-Vi-vs-Doxo-State.csv	Csv

Items per page:

1 - 5 of 61

Publications

Bidirectional communication between nucleotide and substrate binding sites in a type IV multidrug ABC transporter.

Carrillo Victor Hugo Pérez VHP Di Cesare Margot M Rose-Sperling Dania D Javed Waqas W Neuweiler Hannes H Marcoux Julien J Orelle Cédric C Jault Jean-Michel JM Hellmich Ute A UA

Nature communications 20251111 1

ATP-binding cassette (ABC) transporters use ATP to transport substrates across membranes. In type IV ABC transporters, which include many multidrug resistance (MDR) pumps, communication between nucleotide-binding domains (NBDs) and transmembrane domains (TMDs) is mediated via large intracellular domains containing 'coupling helices'. However, how ATP hydrolysis and substrate transport are functionally coordinated remains unclear. In the bacterial type IV MDR transporter BmrA, we identify a conse ...[more]

PMID: 41219214

Similar Datasets

Project description:The identification of multidrug resistant (MDR), extensively and totally drug resistant Mycobacterium tuberculosis (Mtb), in vulnerable sites such as Mumbai, is a grave threat to the control of tuberculosis. The current study aimed at explaining the rapid expression of MDR in Directly Observed Treatment Short Course (DOTS) compliant patients, represents the first study comparing global transcriptional profiles of 3 pairs of clinical Mtb isolates, collected longitudinally at initiation and completion of DOTS. While the isolates were drug susceptible (DS) at onset and MDR at completion of DOTS, they exhibited identical DNA fingerprints at both points of collection. The whole genome transcriptional analysis was performed using total RNA from H37Rv and 3 locally predominant spoligotypes viz. MANU1, CAS and Beijing, hybridized on MTBv3 (BuG@S) microarray, and yielded 36, 98 and 45 differentially expressed genes respectively. Genes encoding transcription factors (sig, rpoB), cell wall biosynthesis (emb genes), protein synthesis (rpl) and additional central metabolic pathways (ppdK, pknH, pfkB) were found to be down regulated in the MDR isolates as compared to the DS isolate of the same genotype. Up regulation of drug efflux pumps, ABC transporters, trans-membrane proteins and stress response transcriptional factors (whiB) in the MDR isolates was observed. The data indicated that Mtb, without specific mutations in drug target genes may persist in the host due to additional mechanisms like drug efflux pumps and lowered rate of metabolism. Furthermore this population of Mtb, which also showed reduced DNA repair activity, would result in selection and stabilization of spontaneous mutations in drug target genes, causing selection of a MDR strain in the presence of drug pressures. Efflux pump such as drrA may play a significant role in increasing fitness of low level drug resistant cells and assist in survival of Mtb till acquisition of drug resistant mutations with least fitness cost. [Data is also available from http://bugs.sgul.ac.uk/E-BUGS-134]