QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

Vol. 15, Issue 10, 4522-4531, October 2004

This Article Full Text Full Text (PDF) All Versions of this Article: E04-06-0445v1 15/10/4522    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Haarer, B. K. Articles by Amberg, D. C. Search for Related Content PubMed PubMed Citation Articles by Haarer, B. K. Articles by Amberg, D. C.

Old Yellow Enzyme Protects the Actin Cytoskeleton from Oxidative Stress

Department of Biochemistry and Molecular Biology, State University of New York, Upstate Medical University, Syracuse, NY 13210

Submitted June 3, 2004; Accepted July 27, 2004
Monitoring Editor: David Drubin

Old Yellow Enzyme (OYE) has long served as a paradigm for the study of flavin-containing NADPH oxido-reductases and yet its physiological role has remained a mystery. A two-hybrid interaction between Oye2p and actin led us to investigate a possible function in the actin cytoskeleton. We found that oye deletion strains have an overly elaborate actin cytoskeleton that cannot be attributed to changes in actin concentration but likely reflect stabilization of actin filaments, resulting in excessive actin assembly. Cells expressing the actin mutant act1-123p, which has a weakened interaction with Oye2p, show comparable defects in actin organization to the oye deletion strain that can be suppressed by overexpression of Oye2p. Similarly, mutation of either conserved cysteine of the potential disulfide pair Cys285-Cys374 in actin completely suppresses the actin organization defect of the oye phenotype. Strains lacking Oye function are also sensitive to oxidative stress as induced by H₂O₂, menadione, and diamide treatment. Mutation of either Cys285 or Cys374 of actin suppresses the sensitivity of oye strains to oxidative stress and in fact confers super-resistance to oxidative stress in otherwise wild-type strains. These results suggest that oxidative damage to actin, like that which has been observed in irreversibly sickled red blood cells, may be a general phenomenon and that OYE functions to control the redox state of actin thereby maintaining the proper plasticity of the actin cytoskeleton. In addition to uncovering a long sought biological function for Old Yellow Enzyme, these results establish that cellular sensitivity to oxidative stress can in part be directly attributed to a specific form (C285-C374 disulfide bond formation) of oxidative damage to actin.

Abbreviations used: AD, activation domain; DBD, DNA binding domain; FMN, flavin mononucleotide; ISC, irreversibly sickled red blood cell; OYE, Old Yellow Enzyme; ROS, reactive oxygen species; RSC, reversibly sickled red blood cell.

^* Corresponding author. E-mail address: ambergd{at}upstate.edu.

This article has been cited by other articles:

				O. Odat, S. Matta, H. Khalil, S. C. Kampranis, R. Pfau, P. N. Tsichlis, and A. M. Makris Old Yellow Enzymes, Highly Homologous FMN Oxidoreductases with Modulating Roles in Oxidative Stress and Programmed Cell Death in Yeast J. Biol. Chem., December 7, 2007; 282(49): 36010 - 36023. [Abstract] [Full Text] [PDF]

				M. E. Farah and D. C. Amberg Conserved Actin Cysteine Residues Are Oxidative Stress Sensors That Can Regulate Cell Death in Yeast Mol. Biol. Cell, April 1, 2007; 18(4): 1359 - 1365. [Abstract] [Full Text] [PDF]

				E. Milgrom, H. Diab, F. Middleton, and P. M. Kane Loss of Vacuolar Proton-translocating ATPase Activity in Yeast Results in Chronic Oxidative Stress J. Biol. Chem., March 9, 2007; 282(10): 7125 - 7136. [Abstract] [Full Text] [PDF]

				D. van den Hemel, A. Brige, S. N. Savvides, and J. Van Beeumen Ligand-induced Conformational Changes in the Capping Subdomain of a Bacterial Old Yellow Enzyme Homologue and Conserved Sequence Fingerprints Provide New Insights into Substrate Binding J. Biol. Chem., September 22, 2006; 281(38): 28152 - 28161. [Abstract] [Full Text] [PDF]

				E. W. Trotter, E. J. Collinson, I. W. Dawes, and C. M. Grant Old Yellow Enzymes Protect against Acrolein Toxicity in the Yeast Saccharomyces cerevisiae. Appl. Envir. Microbiol., July 1, 2006; 72(7): 4885 - 4892. [Abstract] [Full Text] [PDF]

				K.-i. Wakabayashi and S. M. King Modulation of Chlamydomonas reinhardtii flagellar motility by redox poise J. Cell Biol., June 5, 2006; 173(5): 743 - 754. [Abstract] [Full Text] [PDF]

				K. Kitzing, T. B. Fitzpatrick, C. Wilken, J. Sawa, G. P. Bourenkov, P. Macheroux, and T. Clausen The 1.3 A Crystal Structure of the Flavoprotein YqjM Reveals a Novel Class of Old Yellow Enzymes J. Biol. Chem., July 29, 2005; 280(30): 27904 - 27913. [Abstract] [Full Text] [PDF]