Dennis Winge
Professor of Medicine and of Biochemistry
B.A. Concordia College
Ph.D. Duke University
Dennis Winge's Lab Page
Dennis Winge's PubMed Literature Search
Research
Defects in the assembly of the mitochondrial OXPHOS respiratory chain contribute to numerous inherited and acquired diseases of patients with cardiomyopathy, hepatopathy, and neurological disorders. We are interested in elucidating the mechanisms of assembly of respiratory complexes II, III and IV in yeast as an experimental system. Many assembly factors are known that mediate assembly of these respiratory complexes and a large fraction of these are conserved in human cells. Thus, functional studies in yeast contribute to our understanding of the assembly processes in humans.
The first major focus concerns the assembly of the OXPHOS respiratory complexes. The assembly of cytochrome oxidase (CcO) requires the coordinate assembly of subunits translated in both the cytoplasm and mitochondria as well as the insertion of heme and copper redox cofactors. CcO biogenesis commences with the synthesis of Cox1 followed by formation of the heme a cofactor centers and subsequent addition of other subunits. The Cox1 assembly intermediate is conditionally deleterious in cells stalled in subsequent assembly steps. The high spin heme a3 center in Cox1 can generate reactive oxygen species when the Cox1 intermediate accumulates. To avoid the potential danger of this pro-oxidant intermediate, yeast employ chaperone factors to guide this assembly intermediate through the Cox1 maturation process and induce a quality control protease to remove any stalled assembly intermediates. We are focused on elucidating the process of redox cofactor insertion as well as the quality control proteolytic system. We seek to identify additional assembly factors in the CcO biogenesis process and elucidate the mechanism of their assembly function. In these studies, we use a combination of in vitro biochemical, in vivo cellular assays and genetic analyses to elucidate the mechanism and pathway by which these assembly proteins mediate CcO formation. Succinate dehydrogenase (Complex II) and cytochrome c reductase (Complex III) also contain a myriad of metal cofactors including FeS and heme centers. We seek to understand the assembly of these centers. We identified a new factor Mzm1 that mediates a final step in the assembly of Complex III involving the Rieske Fe/S protein. We are also studying the function of two mitochondrial proteins that are important for activation of Complex II.
A second major focus in the Winge laboratory concerns bioavailable pools of copper and zinc within the mitochondrial matrix used in the metallation of metalloproteins. We are using biophysical techniques to structurally characterize the matrix copper and zinc labile complexes and genetic approaches to identify the key transporters that transport copper and zinc ions into the matrix.
The third focus of the group concerns redox chemistry in the mitochondrial intermembrane space (IMS). This compartment is an unusual cellular compartment in that its redox potential is significantly more oxidizing than the matrix or cytoplasm. We are working to understand how redox control modulates CcO biogenesis.
References
1. Winge DR, Robinson NJ (2010) Copper Metallochaperones. Ann Rev Biochem, In Press
2. Atkinson A, Khalimonchuk O, Smith P, Sabic H, Eide D, Winge DR (2010) Role of Mzm1 in Mitochondrial Zinc Maintenance and Respiration. J Biol Chem, In Press
3. Rutter J, Winge DR, Schiffman J (2010) Succinate Dehydrogenase—Assembly, Regulation and Role in Human Disease. Mitochondrion, In Press
4. Bestwick M, Khalimonchuk O, Pierrel F, Winge DR (2010) The role of Coa2 in hemylation of yeast Cox1 revealed by its genetic interaction with Cox10. Mol Cell Biol 30:172-185
5. Khalimonchuk O, Bestwick M, Meunier B, Watts T, Winge DR (2010) Formation of the Redox Cofactor Centers during Cox1 Maturation in Yeast Cytochrome Oxidase. Mol Cell Biol 30:1004-1017
6. Atkinson A, Winge DR (2009) Metal Ion Availability in Mitochondria. Chem Rev 109:4708-21
7. Pierrel F, Khalimonchuk O, Cobine PA, Bestwick M, Winge DR (2008) Coa2 is an assembly factor for yeast cytochrome c oxidase biogenesis facilitating the maturation of Cox1. Mol. Cell Biol. 28:4927-4939
8. Khalimonchuk O, Rigby K, Bestwick M, Pierrel F, Cobine PA, Winge DR (2008) Pet191 is a cytochrome c oxidase assembly factor in Saccharomyces cerevisiae. Eukaryotic Cell 7:1427-31
9. Son M, Leary SC, Romain N, Pierrel F, Winge DR, Haller RG, Elliott JL (2008) Isolated cytochrome c oxidase deficiency in G93A SOD1 mice over-expressing CCS protein. J. Biol. Chem. 283:12267-12275
10. Khalimonchuk O, Winge DR (2008) Function of Cx9C motif proteins within the intermembrane space that are important in the assembly of cytochrome c oxidase. BBA – Mol. Cell Res. 1783:618-628
11. Rigby K, Cobine P, Khalimonchuk O, Winge D (2008) Mapping the interface of the Sco1:Cox2 interaction in cytochrome oxidase biogenesis. J. Biol. Chem. 283(22):15015-22
12. Kumanovics A, Chen O, Li L, Bagley D, Adkins E, Lin H, Dringra N, Outten C, Keller G, Winge D, Ward D, Kaplan J (2008) Identification of FRA1 and FRA2 as genes involved in regulating the yeast iron regulon in response to decreased mitochondrial iron-sulfur cluster synthesis. J. Biol. Chem. 283:10276-10286
13. Pierrel F, Bestwick M, Cobine P, Khalimonchuk O, Cricco J, Winge D (2007) Coa1 links the Mss51 post-translational function to Cox1 cofactor insertion in cytochrome c oxidase assembly. EMBO Journal 26:4335-46
14. Khalimonchuk O, Bird A, Winge D (2007) Identification of a pro-oxidant intermediate in the assembly of cytochrome oxidase. J. Biol. Chem. 282:17442-17449
15. Leary SC, Winge DR (2007) The Janus face of copper: its expanding roles in biology and the pathophysiology of disease. EMBO Reports 8:224-227
16. Coyne HJ, Ciofi-Baffoni S, Banci L, Bertini I, Zhang L, George GN, Winge DR (2007) The characterization and role of zinc binding in yeast Cox4. J. Biol. Chem. 282:8926-8934
17. Rigby K, Zhang L, Cobine PA, George GN, Winge DR (2007) Characterization of the cytochrome c oxidase assembly factor Cox19 of Saccharomyces cerevisiae. J. Biol. Chem. 282:10233-10242
18. Leary SC, Cobine PA, Kaufman BA, Guercin G-H, Mattman A, Palaty J, Lockitch G, Winge DR, Rustin P, Horvath R, Shoubridge EA (2006) The human cytochrome c oxidase assembly proteins SCO1 and SCO2 are involved in the regulation of cellular copper homeostasis. Cell Metabolism 5:9-20
19. Bird AJ, Gordon M, Eide DJ, Winge DR (2006) Repression of ADH1 and ADH3 during zinc deficiency by Zap1-induced intergenic RNA transcripts. EMBO J. 25:5726-5734
20. Pierrel F, Cobine PA, Winge DR (2007) Metal ion availability in mitochondria. Biometals 20(3-4):675-82
Updated 8/15/2010
