Formation of the redox cofactor centers during Cox1 maturation in yeast cytochrome oxidase
Authors: Megan Bestwick
Talina C Watts
Oleh Khalimonchuk
Brigitte Meunier
Dennis R Winge
Journal: Molecular and cellular biology (2010)
Discipline: MOLECULAR BIOLOGY & GENETICS
Keywords: Cytochrome c oxidase; Cox1; redox cofactors; heme a center; heterobimetallic
Affiliations: University of Utah Health Sciences Center, Department of Medicine, Salt Lake City, UT 84132, USA., University of Utah Health Sciences Center, Departments of Medicine and Biochemistry, Salt Lake City, Utah 84132, Centre de Génétique Moléculaire, CNRS, Avenue de la Terrasse, 91198 Gif-sur-Yvette, France

Abstract:  The biogenesis of cytochrome c oxidase initiates with synthesis and maturation of the mitochondrion-encoded Cox1 subunit prior to the addition of other subunits. Cox1 contains redox cofactors, including the low-spin heme a center and the heterobimetallic heme a(3):Cu(B) center. We sought to identify the step in the maturation of Cox1 in which the redox cofactor centers are assembled. Newly synthesized Cox1 is incorporated within one early assembly intermediate containing Mss51 in Saccharomyces cerevisiae. Subsequent Cox1 maturation involves the progression to downstream assembly intermediates involving Coa1 and Shy1. We show that the two heme a cofactor sites in Cox1 form downstream of Mss51- and Coa1-containing Cox1 intermediates. These Cox1 intermediates form normally in cells defective in heme a biosynthesis or in cox1 mutant strains with heme a axial His mutations. In contrast, the Shy1-containing Cox1 assembly intermediate is perturbed in the absence of heme a. Heme a(3) center formation in Cox1 appears to be chaperoned by Shy1. Cu(B) site formation occurs near or at the Shy1-containing Cox1 assembly intermediate also. The Cu(B) metallochaperone Cox11 transiently interacts with Shy1 by coimmunoprecipitation. The Shy1-containing Cox1 complex is markedly attenuated in cells lacking Cox11 but is partially restored with a nonfunctional Cox11 mutant. Thus, formation of the heterobimetallic Cu(B):heme a(3) site likely occurs in the Shy1-containing Cox1 complex. 

Retracted: No

PubPeer comments:
 May 2017 Correction, found here: https: mcb.asm.org content 37 11 e00127-17.long It reads:  * Fig. 1D: The anti-F1 control immunoblot was inadvertently duplicated. The correct anti-F1 blot from the original experiment is shown below. This is the monomeric form of mitochondrial F1F0 ATPase (complex V) as a loading control. * Fig. 1E: The anti-F1 immunoblot was inadvertently a duplicate of the control blot in Fig. 5A. Therefore, we redid the Fig. 1E study using the original samples from 2009. * Fig. 2C, lanes 5 and 6: The anti-F1 immunoblot was a duplicate of the blot in Fig. 1D. The original samples were rerun, and the correct anti-Myc and anti-F1 blots are shown below. * Fig. 5A, lanes 3 and 4: The anti-F1 blot was a duplicate of the control in Fig. 1E, as mentioned above. We reran the original samples, and the correct anti-Myc and anti-F1 blots for Coa1-Myc samples from wild-type (WT) and cox10u0394 cells are shown below. We regret these errors, but they do not affect the interpretation of the experiments and the conclusions of the study. , 
 Here are the original Figures 1, 2C, and 5A, with the duplications marked in red and blue, as reported to the journal in September 2016. ![file] This paper was part of a structured search for image duplications in Molecular and Cellular Biology paper, which was published here: https: mcb.asm.org content 38 20 e00309-18.long ,