PMID- 33662401
OWN - NLM
STAT- MEDLINE
DCOM- 20210909
LR  - 20220226
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 296
DP  - 2021 Jan-Jun
TI  - A genome-wide copper-sensitized screen identifies novel regulators of 
      mitochondrial cytochrome c oxidase activity.
PG  - 100485
LID - S0021-9258(21)00259-3 [pii]
LID - 10.1016/j.jbc.2021.100485 [doi]
LID - 100485
AB  - Copper is essential for the activity and stability of cytochrome c oxidase (CcO), 
      the terminal enzyme of the mitochondrial respiratory chain. Loss-of-function 
      mutations in genes required for copper transport to CcO result in fatal human 
      disorders. Despite the fundamental importance of copper in mitochondrial and 
      organismal physiology, systematic identification of genes that regulate 
      mitochondrial copper homeostasis is lacking. To discover these genes, we 
      performed a genome-wide screen using a library of DNA-barcoded yeast deletion 
      mutants grown in copper-supplemented media. Our screen recovered a number of 
      genes known to be involved in cellular copper homeostasis as well as genes 
      previously not linked to mitochondrial copper biology. These newly identified 
      genes include the subunits of the adaptor protein 3 complex (AP-3) and components 
      of the cellular pH-sensing pathway Rim20 and Rim21, both of which are known to 
      affect vacuolar function. We find that AP-3 and Rim mutants exhibit decreased 
      vacuolar acidity, which in turn perturbs mitochondrial copper homeostasis and CcO 
      function. CcO activity of these mutants could be rescued by either restoring 
      vacuolar pH or supplementing growth media with additional copper. Consistent with 
      these genetic data, pharmacological inhibition of the vacuolar proton pump leads 
      to decreased mitochondrial copper content and a concomitant decrease in CcO 
      abundance and activity. Taken together, our study uncovered novel genetic 
      regulators of mitochondrial copper homeostasis and provided a mechanism by which 
      vacuolar pH impacts mitochondrial respiration through copper homeostasis.
CI  - Copyright (c) 2021 The Authors. Published by Elsevier Inc. All rights reserved.
FAU - Garza, Natalie M
AU  - Garza NM
AD  - Department of Biochemistry and Biophysics, Texas A&M University, College Station, 
      Texas, USA.
FAU - Griffin, Aaron T
AU  - Griffin AT
AD  - Department of Biochemistry and Biophysics, Texas A&M University, College Station, 
      Texas, USA.
FAU - Zulkifli, Mohammad
AU  - Zulkifli M
AD  - Department of Biochemistry and Biophysics, Texas A&M University, College Station, 
      Texas, USA.
FAU - Qiu, Chenxi
AU  - Qiu C
AD  - Department of Biochemistry and Biophysics, Texas A&M University, College Station, 
      Texas, USA.
FAU - Kaplan, Craig D
AU  - Kaplan CD
AD  - Department of Biochemistry and Biophysics, Texas A&M University, College Station, 
      Texas, USA.
FAU - Gohil, Vishal M
AU  - Gohil VM
AD  - Department of Biochemistry and Biophysics, Texas A&M University, College Station, 
      Texas, USA. Electronic address: vgohil@tamu.edu.
LA  - eng
GR  - F31 GM128339/GM/NIGMS NIH HHS/United States
GR  - R01 GM097260/GM/NIGMS NIH HHS/United States
GR  - R01 GM111672/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20210301
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Culture Media)
RN  - 0 (Saccharomyces cerevisiae Proteins)
RN  - 789U1901C5 (Copper)
RN  - EC 1.9.3.1 (Electron Transport Complex IV)
SB  - IM
MH  - Copper/*metabolism
MH  - Culture Media
MH  - Electron Transport Complex IV/genetics/*metabolism
MH  - Genome, Fungal
MH  - High-Throughput Nucleotide Sequencing/methods
MH  - Homeostasis
MH  - Mitochondria/*enzymology
MH  - Saccharomyces cerevisiae/enzymology/genetics/growth & development/*metabolism
MH  - Saccharomyces cerevisiae Proteins/genetics/*metabolism
MH  - Sequence Deletion
PMC - PMC8027276
OTO - NOTNLM
OT  - AP-3
OT  - copper
OT  - cytochrome c oxidase
OT  - mitochondria
OT  - pH
OT  - rim20
OT  - rim21
OT  - vacuole
COIS- Conflict of interest The authors declare that they have no conflicts of interest 
      with the contents of this article.
EDAT- 2021/03/05 06:00
MHDA- 2021/09/10 06:00
PMCR- 2021/03/01
CRDT- 2021/03/04 20:12
PHST- 2020/12/30 00:00 [received]
PHST- 2021/02/22 00:00 [revised]
PHST- 2021/02/25 00:00 [accepted]
PHST- 2021/03/05 06:00 [pubmed]
PHST- 2021/09/10 06:00 [medline]
PHST- 2021/03/04 20:12 [entrez]
PHST- 2021/03/01 00:00 [pmc-release]
AID - S0021-9258(21)00259-3 [pii]
AID - 100485 [pii]
AID - 10.1016/j.jbc.2021.100485 [doi]
PST - ppublish
SO  - J Biol Chem. 2021 Jan-Jun;296:100485. doi: 10.1016/j.jbc.2021.100485. Epub 2021 
      Mar 1.