Structurally dependent redox property of ribonucleotide reductase subunit p53R2

Lijun Xue, Bingsen Zhou, Xiyong Liu, Tieli Wang, Jennifer Shih, Christina Qi, Yvonne Heung, Yun Yen

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

p53R2 is a newly identified small subunit of ribonucleotide reductase (RB) and plays a key role in supplying precursors for DNA repair in a p53-dependent manner. Currently, we are studying the redox property, structure, and function of p53R2. In cell-free systems, p53R2 did not oxidize a reactive oxygen species (ROS) indicator carboxy-H2DCFDA, but another class I RR small subunit, hRRM2, did. Further studies showed that purified recombinant p53R2 protein has catalase activity, which breaks down H2O2. Overexpression of p53R2 reduced intracellular ROS and protected the mitochondrial membrane potential against oxidative stress, whereas overexpression of hRRM2 did not and resulted in a collapse of mitochondrial membrane potential. In a site-directed mutagenesis study, antioxidant activity was abrogated in p53R2 mutants Y331F, Y285F, Y49F, and Y241H, but not Y164F or Y164C. The fluorescence intensity in mutants oxidizing carboxy-H 2DCFDA, in order from highest to lowest, was Y331F > Y285F > Y49F > Y241H > wild-type p53R2. This indicates that Y331, Y285, Y49, and Y241 in p53R2 are critical residues involved in scavenging ROS. Of interest, the ability to oxidize carboxy-H2DCFDA indicated by fluorescence intensity was negatively correlated with RR activity from wild-type p53R2, mutants Y331F, Y285F, and Y49F. Our findings suggest that p53R2 may play a key role in defending oxidative stress by scavenging ROS, and this antioxidant property is also important for its fundamental enzymatic activity.

Original languageEnglish
Pages (from-to)1900-1905
Number of pages6
JournalCancer Research
Volume66
Issue number4
DOIs
Publication statusPublished - Feb 15 2006
Externally publishedYes

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Ribonucleotide Reductases
Oxidation-Reduction
Reactive Oxygen Species
Mitochondrial Membrane Potential
Oxidative Stress
Antioxidants
Fluorescence
Cell-Free System
Site-Directed Mutagenesis
Recombinant Proteins
DNA Repair
Catalase
2',7'-dichlorodihydrofluorescein diacetate

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Structurally dependent redox property of ribonucleotide reductase subunit p53R2. / Xue, Lijun; Zhou, Bingsen; Liu, Xiyong; Wang, Tieli; Shih, Jennifer; Qi, Christina; Heung, Yvonne; Yen, Yun.

In: Cancer Research, Vol. 66, No. 4, 15.02.2006, p. 1900-1905.

Research output: Contribution to journalArticle

Xue, L, Zhou, B, Liu, X, Wang, T, Shih, J, Qi, C, Heung, Y & Yen, Y 2006, 'Structurally dependent redox property of ribonucleotide reductase subunit p53R2', Cancer Research, vol. 66, no. 4, pp. 1900-1905. https://doi.org/10.1158/0008-5472.CAN-05-2656
Xue, Lijun ; Zhou, Bingsen ; Liu, Xiyong ; Wang, Tieli ; Shih, Jennifer ; Qi, Christina ; Heung, Yvonne ; Yen, Yun. / Structurally dependent redox property of ribonucleotide reductase subunit p53R2. In: Cancer Research. 2006 ; Vol. 66, No. 4. pp. 1900-1905.
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