PUBLICATION

PYCR1 and PYCR2 Interact and Collaborate with RRM2B to Protect Cells from Overt Oxidative Stress

Authors
Kuo, M.L., Lee, M.B., Tang, M., den Besten, W., Hu, S., Sweredoski, M.J., Hess, S., Chou, C.M., Changou, C.A., Su, M., Jia, W., Su, L., Yen, Y.
ID
ZDB-PUB-170214-54
Date
2016
Source
Scientific Reports   6: 18846 (Journal)
Registered Authors
Keywords
Cell signalling, Protein–protein interaction networks
MeSH Terms
  • Animals
  • Antioxidants/metabolism
  • Cell Cycle Checkpoints
  • Cell Cycle Proteins/genetics
  • Cell Cycle Proteins/metabolism*
  • Cell Line
  • Gene Knockdown Techniques
  • Gene Silencing
  • Humans
  • Isoenzymes
  • Mass Spectrometry/methods
  • Mitochondria/genetics
  • Mitochondria/metabolism
  • Multiprotein Complexes/metabolism
  • Oxidative Stress*
  • Protein Binding
  • Protein Interaction Mapping
  • Protein Transport
  • Pyrroline Carboxylate Reductases/genetics
  • Pyrroline Carboxylate Reductases/metabolism*
  • Recombinant Fusion Proteins
  • Ribonucleotide Reductases/genetics
  • Ribonucleotide Reductases/metabolism*
  • Signal Transduction
  • Telomerase/genetics
  • Telomerase/metabolism
  • Tumor Suppressor Protein p53/metabolism
  • Zebrafish
PubMed
26733354 Full text @ Sci. Rep.
Abstract
Ribonucleotide reductase small subunit B (RRM2B) is a stress response protein that protects normal human fibroblasts from oxidative stress. However, the underlying mechanism that governs this function is not entirely understood. To identify factors that interact with RRM2B and mediate anti-oxidation function, large-scale purification of human Flag-tagged RRM2B complexes was performed. Pyrroline-5-carboxylate reductase 1 and 2 (PYCR1, PYCR2) were identified by mass spectrometry analysis as components of RRM2B complexes. Silencing of both PYCR1 and PYCR2 by expressing short hairpin RNAs induced defects in cell proliferation, partial fragmentation of the mitochondrial network, and hypersensitivity to oxidative stress in hTERT-immortalized human foreskin fibroblasts (HFF-hTERT). Moderate overexpression of RRM2B, comparable to stress-induced level, protected cells from oxidative stress. Silencing of both PYCR1 and PYCR2 completely abolished anti-oxidation activity of RRM2B, demonstrating a functional collaboration of these metabolic enzymes in response to oxidative stress.
Genes / Markers
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Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping