Flexible Target Recognition of the Intrinsically Disordered DNA-Binding Domain of CytR Monitored by Single-Molecule Fluorescence Spectroscopy

Overview

Journal J Phys Chem B

Publisher American Chemical Society

Specialty Chemistry

Date 2022 Aug 15

PMID 35969476

Authors

Shrutarshi Mitra

Hiroyuki Oikawa

Divya Rajendran

Toshiyuki Kowada

Shin Mizukami

Athi N Naganathan

Satoshi Takahashi

Affiliations

Soon will be listed here.

Abstract

The intrinsically disordered DNA-binding domain of cytidine repressor (CytR-DBD) folds in the presence of target DNA and regulates the expression of multiple genes in . To explore the conformational rearrangements in the unbound state and the target recognition mechanisms of CytR-DBD, we carried out single-molecule Förster resonance energy transfer (smFRET) measurements. The smFRET data of CytR-DBD in the absence of DNA show one major and one minor population assignable to an expanded unfolded state and a compact folded state, respectively. The population of the folded state increases and decreases upon titration with salt and denaturant, respectively, in an apparent two-state manner. The peak FRET efficiencies of both the unfolded and folded states change continuously with denaturant concentration, demonstrating the intrinsic flexibility of the DNA-binding domain and the deviation from a strict two-state transition. Remarkably, the CytR-DBD exhibits a compact structure when bound to both the specific and nonspecific DNA; however, the peak FRET efficiencies of the two structures are slightly but consistently different. The observed conformational heterogeneity highlights the potential structural changes required for CytR to bind variably spaced operator sequences.

Citing Articles

Functional regulation of an intrinsically disordered protein via a conformationally excited state.

Madhurima K, Nandi B, Munshi S, Naganathan A, Sekhar A Sci Adv. 2023; 9(26):eadh4591.

PMID: 37379390 PMC: 10306299. DOI: 10.1126/sciadv.adh4591.

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