The Human RecQ4 Helicase Contains a Functional RecQ C-terminal Region (RQC) That Is Essential for Activity

Mojumdar, Aditya; March, Matteo De; Marino, Francesca; Onesti, Silvia

doi:10.1074/jbc.m116.767954

Cited by 17 publications

(19 citation statements)

References 38 publications

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“…It provides information regarding the sizes and shapes of proteins and complexes over a broad range of molecular sizes ranging from several kDa to GDa [32][33][34]. In particular, SAXS can comprehensively characterize dynamic processes in systems where macromolecular structures are evolving under varying experimental conditions (e.g., time, pH, pressure, or different reagents) [35][36][37][38]. In general, a highly purified monodisperse ideal sample without intermolecular interactions is essential for the reconstruction of 3D models from SAXS data.…”

Section: Introductionmentioning

confidence: 99%

Chemically Denatured Structures of Porcine Pepsin using Small-Angle X-ray Scattering

et al. 2019

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Porcine pepsin is a gastric aspartic proteinase that reportedly plays a pivotal role in the digestive process of many vertebrates. We have investigated the three-dimensional (3D) structure and conformational transition of porcine pepsin in solution over a wide range of denaturant urea concentrations (0–10 M) using Raman spectroscopy and small-angle X-ray scattering. Furthermore, 3D GASBOR ab initio structural models, which provide an adequate conformational description of pepsin under varying denatured conditions, were successfully constructed. It was shown that pepsin molecules retain native conformation at 0–5 M urea, undergo partial denaturation at 6 M urea, and display a strongly unfolded conformation at 7–10 M urea. According to the resulting GASBOR solution models, we identified an intermediate pepsin conformation that was dominant during the early stage of denaturation. We believe that the structural evidence presented here provides useful insights into the relationship between enzymatic activity and conformation of porcine pepsin at different states of denaturation.

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Section: Introductionmentioning

confidence: 99%

Chemically Denatured Structures of Porcine Pepsin using Small-Angle X-ray Scattering

et al. 2019

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“…The amino acid residues shown to play a role in enzyme function33, were mutated to alanine (shown in Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

DNA-conjugated gold nanoparticles based colorimetric assay to assess helicase activity: a novel route to screen potential helicase inhibitors

Deka

Mojumdar

Parisse

et al. 2017

Sci Rep

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Helicase are essential enzymes which are widespread in all life-forms. Due to their central role in nucleic acid metabolism, they are emerging as important targets for anti-viral, antibacterial and anti-cancer drugs. The development of easy, cheap, fast and robust biochemical assays to measure helicase activity, overcoming the limitations of the current methods, is a pre-requisite for the discovery of helicase inhibitors through high-throughput screenings. We have developed a method which exploits the optical properties of DNA-conjugated gold nanoparticles (AuNP) and meets the required criteria. The method was tested with the catalytic domain of the human RecQ4 helicase and compared with a conventional FRET-based assay. The AuNP-based assay produced similar results but is simpler, more robust and cheaper than FRET. Therefore, our nanotechnology-based platform shows the potential to provide a useful alternative to the existing conventional methods for following helicase activity and to screen small-molecule libraries as potential helicase inhibitors.

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“…During the preparation of this manuscript a study by Mojumdar et al 48. was published, which suggested that RecQ4 features a functional RQC domain that is essential for RecQ4 activity.…”

Section: Discussionmentioning

confidence: 99%

The structural and functional characterization of human RecQ4 reveals insights into its helicase mechanism

Kaiser¹,

Sauer²,

Kisker³

2017

Nat Commun

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RecQ4 is a member of the RecQ helicase family, an evolutionarily conserved class of enzymes, dedicated to preserving genomic integrity by operating in telomere maintenance, DNA repair and replication. While reduced RecQ4 activity is associated with cancer predisposition and premature aging, RecQ4 upregulation is related to carcinogenesis and metastasis. Within the RecQ family, RecQ4 assumes an exceptional position, lacking several characteristic RecQ domains. Here we present the crystal structure of human RecQ4, encompassing the conserved ATPase core and a novel C-terminal domain that lacks resemblance to the RQC domain observed in other RecQ helicases. The new domain features a zinc-binding site and two distinct types of winged-helix domains, which are not involved in canonical DNA binding or helicase activity. Based on our structural and functional analysis, we propose that RecQ4 exerts a helicase mechanism, which may be more closely related to bacterial RecQ helicases than to its human family members.

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The Human RecQ4 Helicase Contains a Functional RecQ C-terminal Region (RQC) That Is Essential for Activity

Cited by 17 publications

References 38 publications

Chemically Denatured Structures of Porcine Pepsin using Small-Angle X-ray Scattering

Chemically Denatured Structures of Porcine Pepsin using Small-Angle X-ray Scattering

DNA-conjugated gold nanoparticles based colorimetric assay to assess helicase activity: a novel route to screen potential helicase inhibitors

The structural and functional characterization of human RecQ4 reveals insights into its helicase mechanism

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