A Fluorimetric Readout Reporting the Kinetics of Nucleotide-induced Human Ribonucleotide Reductase Oligomerization

Overview

Journal Chembiochem

Specialty Biochemistry

Date 2014 Sep 27

PMID 25256246

Citations 7

Authors

Yuan Fu

Hongyu Lin

Somsinee Wisitpitthaya

William A Blessing

Yimon Aye

Affiliations

Soon will be listed here.

Abstract

Human ribonucleotide reductase (hRNR) is a target of nucleotide chemotherapeutics in clinical use. The nucleotide-induced oligomeric regulation of hRNR subunit α is increasingly being recognized as an innate and drug-relevant mechanism for enzyme activity modulation. In the presence of negative feedback inhibitor dATP and leukemia drug clofarabine nucleotides, hRNR-α assembles into catalytically inert hexameric complexes, whereas nucleotide effectors that govern substrate specificity typically trigger α-dimerization. Currently, both knowledge of and tools to interrogate the oligomeric assembly pathway of RNR in any species in real time are lacking. We therefore developed a fluorimetric assay that reliably reports on oligomeric state changes of α with high sensitivity. The oligomerization-directed fluorescence quenching of hRNR-α, covalently labeled with two fluorophores, allows for direct readout of hRNR dimeric and hexameric states. We applied the newly developed platform to reveal the timescales of α self-assembly, driven by the feedback regulator dATP. This information is currently unavailable, despite the pharmaceutical relevance of hRNR oligomeric regulation.

Citing Articles

Still no Rest for the Reductases: Ribonucleotide Reductase (RNR) Structure and Function: An Update.

Long M, Ly P, Aye Y Subcell Biochem. 2022; 99:155-197.

PMID: 36151376 DOI: 10.1007/978-3-031-00793-4_5.

Clofarabine Commandeers the RNR-α-ZRANB3 Nuclear Signaling Axis.

Long M, Zhao Y, Aye Y Cell Chem Biol. 2019; 27(1):122-133.e5.

PMID: 31836351 PMC: 6980387. DOI: 10.1016/j.chembiol.2019.11.012.

The more the merrier: how homo-oligomerization alters the interactome and function of ribonucleotide reductase.

Long M, Van Hall-Beauvais A, Aye Y Curr Opin Chem Biol. 2019; 54:10-18.

PMID: 31734537 PMC: 7131891. DOI: 10.1016/j.cbpa.2019.09.003.

[Silencing RRM1 gene reverses paclitaxel resistance in human breast cancer cell line MCF- 7/R by inducing cell apoptosis].

Tian N, Zhou L, Yang D, Wu H, Ma Y, Lu L Nan Fang Yi Ke Da Xue Xue Bao. 2019; 39(3):304-312.

PMID: 31068300 PMC: 6765681. DOI: 10.12122/j.issn.1673-4254.2019.03.08.

Nuclear RNR-α antagonizes cell proliferation by directly inhibiting ZRANB3.

Fu Y, Long M, Wisitpitthaya S, Inayat H, Pierpont T, Elsaid I Nat Chem Biol. 2018; 14(10):943-954.

PMID: 30150681 PMC: 6171530. DOI: 10.1038/s41589-018-0113-5.

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