title: PYCR1 and PYCR2 Interact and Collaborate with RRM2B to Protect Cells from Overt Oxidative Stress.
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.
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute at City of Hope, Duarte, CA 91010, USA.
authors.forename: Mei-Ling
authors.initials: ML
authors.lastname: Kuo
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute at City of Hope, Duarte, CA 91010, USA.
authors.forename: Mabel Bin-Er
authors.initials: MB
authors.lastname: Lee
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute at City of Hope, Duarte, CA 91010, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Tang
authors.affiliation: Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA.
authors.forename: Willem
authors.initials: W
authors.lastname: den Besten
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute at City of Hope, Duarte, CA 91010, USA.
authors.forename: Shuya
authors.initials: S
authors.lastname: Hu
authors.affiliation: Proteome Exploration Laboratory, Division of Biology and Biological Engineering, Beckman Institute, California Institute of Technology, Pasadena, CA, 91125.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Sweredoski
authors.affiliation: Proteome Exploration Laboratory, Division of Biology and Biological Engineering, Beckman Institute, California Institute of Technology, Pasadena, CA, 91125.
authors.forename: Sonja
authors.initials: S
authors.lastname: Hess
authors.affiliation: Department of Biochemistry and Molecular Cell Biology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 110.
authors.forename: Chih-Ming
authors.initials: CM
authors.lastname: Chou
authors.affiliation: Integrated Laboratory, Center of Translational Medicine, Taipei Medical University, Taipei, Taiwan 110.|Graduate Institute of Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan 110.|Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan 110.
authors.forename: Chun A
authors.initials: CA
authors.lastname: Changou
authors.affiliation: University of Hawaii Cancer Center, HI 96813, USA.
authors.forename: Mingming
authors.initials: M
authors.lastname: Su
authors.affiliation: University of Hawaii Cancer Center, HI 96813, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Jia
authors.affiliation: Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan 110.
authors.forename: Leila
authors.initials: L
authors.lastname: Su
authors.affiliation: Department of Molecular Pharmacology, Beckman Research Institute at City of Hope, Duarte, CA 91010, USA.|Department of Biochemistry and Molecular Cell Biology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan 110.|Integrated Laboratory, Center of Translational Medicine, Taipei Medical University, Taipei, Taiwan 110.|Graduate Institute of Translational Medicine, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan 110.|Graduate Institute of Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan 110.
authors.forename: Yun
authors.initials: Y
authors.lastname: Yen
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D059447:Cell Cycle Checkpoints; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D055785:Gene Knockdown Techniques; D020868:Gene Silencing; D006801:Humans; D007527:Isoenzymes; D013058:Mass Spectrometry / Q000379:methods; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism; D046912:Multiprotein Complexes / Q000378:metabolism; D018384:Oxidative Stress; D011485:Protein Binding; D025941:Protein Interaction Mapping; D021381:Protein Transport; D011762:Pyrroline Carboxylate Reductases / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012264:Ribonucleotide Reductases / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D015027:Zebrafish; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase
chemical_list: D000975:Antioxidants; D018797:Cell Cycle Proteins; D007527:Isoenzymes; D046912:Multiprotein Complexes; D011762:Pyrroline Carboxylate Reductases; D011993:Recombinant Fusion Proteins; D012264:Ribonucleotide Reductases; D019098:Telomerase; D016159:Tumor Suppressor Protein p53; D000097502:delta-1-Pyrroline-5-Carboxylate Reductase; C405947:RRM2B protein, human