| 2010 |
ChChd3 (CHCHD3/Mic19) is a peripheral protein of the mitochondrial inner membrane facing the intermembrane space. RNAi knockdown in HeLa cells caused mitochondrial fragmentation, reduced OPA1 protein levels, impaired fusion, and aberrant cristae with reduced crista junction opening diameter (~50% reduction). ChChd3 interacts with inner membrane proteins mitofilin and OPA1, and outer membrane protein Sam50; knockdown led to near-complete loss of mitofilin and Sam50, establishing ChChd3 as a scaffolding protein stabilizing complexes that maintain crista architecture and protein import. |
RNAi knockdown in HeLa cells, Co-IP/binding partner analysis, ultrastructural analysis (electron microscopy), oxygen consumption and glycolytic rate measurements |
The Journal of biological chemistry |
High |
21081504
|
| 2007 |
CHCHD3 (ChChd3) was identified as a novel substrate of cAMP-dependent protein kinase (PKA) using an analog-sensitive PKA catalytic subunit (M120G mutant) with bulky N6-substituted ATP analogs, establishing PKA as a writer of CHCHD3 phosphorylation. |
Chemical genetics approach with analog-sensitive PKA catalytic subunit, in vitro kinase assay with N6-substituted ATP analogs, mass spectrometry substrate identification |
The Journal of biological chemistry |
Medium |
17242405
|
| 2012 |
CHCHD3 import into the mitochondrial IMS requires both N-terminal myristoylation and the C-terminal CHCH domain. Myristoylation promotes binding to the outer membrane; the CHCH domain mediates translocation across the outer membrane. The disulfide relay via Mia40 occurs preferentially between Cys193 (second cysteine in helix 1) and Mia40 Cys55. Each of the four CHCH domain cysteines is essential for protein folding and binding to mitofilin and Sam50, but not for import per se. |
Cysteine mutagenesis, myristoylation site mutagenesis (G2A), in vitro import assays, binding assays to Mia40/mitofilin/Sam50, subcellular fractionation |
The Journal of biological chemistry |
High |
23019327
|
| 2015 |
MIC19 (CHCHD3) undergoes oxidation in mitochondria and requires the MIA pathway for mitochondrial localization. Yeast Mic19 exists in two redox forms; the intramolecular disulfide-bonded form is specifically bound to Mic60 of the MICOS complex. Mic19 oxidation is not essential for MICOS integration but promotes MICOS assembly and proper inner membrane morphology. |
Redox state analysis (non-reducing SDS-PAGE), MIA pathway mutant analysis, Co-IP (Mic19-Mic60 interaction), yeast genetics, immunofluorescence |
Molecular and cellular biology |
High |
26416881
|
| 2017 |
Using genetically encoded tags (miniSOG and APEX2) and electron tomography, Mic19 (CHCHD3) was localized at nanoscale resolution to crista junctions, distributed in a network along the mitochondrial periphery, and enriched inside cristae in cardiac and astrocyte cell lines. Mic19 was found associated with cytochrome c oxidase subunit IV at crista junctions. |
Genetic tagging with miniSOG and APEX2, electron tomography, subcellular fractionation |
Journal of cell science |
High |
28808085
|
| 2018 |
MIC19 (CHCHD3) is N-myristoylated at its N-terminus. In vitro and in vivo metabolic labeling confirmed N-myristoylation. G2A (non-myristoylated) mutant analysis showed that myristoylation is required for proper mitochondrial targeting and membrane binding of MIC19. Additionally, myristoylation of MIC19 is required for the protein-protein interaction between MIC19 and SAMM50. |
In vitro and in vivo metabolic labeling with myristate, G2A mutagenesis, immunofluorescence, subcellular fractionation, co-immunoprecipitation |
PloS one |
High |
30427857
|
| 2019 |
Mic19 (CHCHD3) directly interacts with outer membrane protein Sam50 and inner membrane protein Mic60 to form the Sam50-Mic19-Mic60 axis, which connects SAM and MICOS complexes to assemble the MIB supercomplex and mediates mitochondrial outer-inner membrane contact. OMA1 protease cleaves Mic19 at its N-terminus under physiological stress, disrupting this axis and causing loss of crista junctions, abnormal cristae distribution, and reduced ATP production. Sam50 acts as an anchoring point at the outer membrane guiding crista junction formation. |
Co-IP, OMA1-mediated cleavage assays, MIB supercomplex analysis, ATP production measurements, mitochondrial morphology analysis, electron microscopy of cristae |
Cell death and differentiation |
High |
31097788
|
| 2023 |
Fasting induces MIC19 (CHCHD3) upregulation in mouse liver, promoting cristae formation. Enforced hepatic MIC19 expression promotes mitochondrial respiration, fatty acid oxidation, and suppresses gluconeogenesis. MIC19-driven cristae formation increases uridine phosphorylase UPP2 activity and uracil accumulation, which signals to promote locomotion. |
Comparative mouse proteomics, hepatic MIC19 transgenic overexpression, Seahorse respirometry, metabolite profiling, dietary uracil supplementation experiments |
Cell metabolism |
Medium |
37473754
|
| 2024 |
Mic19 (CHCHD3) regulates ER-mitochondria contacts through the EMC2-SLC25A46-Mic19 axis. Liver-specific Mic19 knockout in mice leads to reduction of ER-mitochondrial contacts, mitochondrial lipid metabolism disorder, disorganization of mitochondrial cristae, and mitochondrial unfolded protein stress response in hepatocytes, impairing fatty acid β-oxidation and spontaneously triggering NASH and liver fibrosis. Re-expression of Mic19 in LKO hepatocytes rescued liver disease. |
Liver-specific conditional knockout (LKO), ER-mitochondria contact site quantification, mitochondrial fractionation, fatty acid oxidation assays, hepatic rescue re-expression experiments, in vivo mouse models |
Nature communications |
High |
38168065
|
| 2024 |
ASB1 E3 ubiquitin ligase interacts with CHCHD3 and promotes its degradation via K48-linked ubiquitination. Loss of ASB1 stabilizes CHCHD3, which activates ROS signaling to promote prostate cancer cell proliferation, clonogenicity, and migration. |
Quantitative mass spectrometry interactome analysis, co-immunoprecipitation, cycloheximide chase assay, ubiquitination assay, cell rescue experiments |
American journal of cancer research |
Medium |
39113857
|
| 2025 |
Under hypoxic conditions, HIF-1α binds the USP3 promoter to upregulate USP3 expression, which in turn stabilizes MIC19 (CHCHD3) through K48-linked deubiquitination, preventing its proteasomal degradation and promoting NSCLC progression. |
ChIP assay (HIF-1α at USP3 promoter), ubiquitination assay, co-immunoprecipitation (USP3-MIC19), in vitro cell proliferation/invasion assays, in vivo xenograft mouse model |
Acta pharmacologica Sinica |
Medium |
40770539
|
| 2026 |
SLC25A6 directly interacts with MIC60 and competitively inhibits MIC19 (CHCHD3) binding to MIC60, disrupting the MICOS complex. A SLC25A6 T126A mutant failed to bind MIC60, abrogating its ability to destabilize MICOS and cause mitofission. This mechanistically links metabolic stress to mitochondrial fragmentation via displacement of MIC19 from MIC60. |
Co-immunoprecipitation (SLC25A6-MIC60, competitive with MIC19), site-directed mutagenesis (T126A), mitochondrial morphology analysis, apoptosis assays |
Cell death & disease |
Medium |
42020360
|
| 2026 |
CHCHD3 knockdown in lung adenocarcinoma cells impaired mitochondrial energy metabolism and caused excessive ROS production. IP-MS and Co-IP validated SAMM50 and VDAC1/2 as direct CHCHD3 binding partners, with disruption of these interactions linked to ROS accumulation. |
IP-MS, co-immunoprecipitation, Seahorse metabolic analysis, ROS measurement, cell cycle analysis, apoptosis assays |
Anti-cancer agents in medicinal chemistry |
Low |
42261148
|