Tom70-based transcriptional regulation of mitochondrial biogenesis and aging
- Buck Institute for Research on Aging, United States
- Healthy Longevity Programme, Yong Loo Lin School of Medicine, National University of Singapore, Singapore
- Centre for Healthy Longevity, National University Health System, Singapore
- Singapore Institute of Clinical Sciences, A(∗)STAR, Singapore
- USC Leonard Davis School of Gerontology, University of Southern California, United States
Figures
Figure 1 with 1 supplement
Tom70 regulates the transcriptional activity of mitochondrial proteins.
(A) Representative images for proteins localized to different sub-compartments of mitochondria in cells overexpressing (OE) different TOM proteins and HAP4 from pGal promoter. HAP4 OE increased the …
Figure 1—figure supplement 1
Additional data demonstrating that Tom70 regulates the transcription of mitochondrial proteins.
(A) Quantification of Figure 1A. Dash line boxes indicate the ones with p < 0.05 from unpaired two-tailed t test. (B) Representative images for proteins localized to different sub-compartments of …
Figure 2 with 1 supplement
The transcription factors involved in the Tom70-dependent transcriptional regulation of mitochondrial proteins.
(A) mRNA abundance of different transcription factors (TF) quantified by RT-qPCR in TOM70 OE strain and normalized to control cells. All different yeast strains, including wild-type control, were …
Figure 2—figure supplement 1
ROS is partially required for the mitochondrial biogenesis program downstream of TOM70 OE.
GFP signal of mitochondrial proteins were quantified in different strains and normalized to wild type control. Mitochondrial proteins were visualized by endogenous C-terminal GFP tagging and …
Figure 3 with 1 supplement
Tom70 regulates the abundance of mtDNA.
(A, B) Representative images and quantification of mtDNA in wild type and ∆tom70 cells were stained by Hoechst dye. Yellow arrowheads point to the mtDNA. Bar graph are the mean and s.e.m. from 87 …
Figure 3—figure supplement 1
The role of mtDNA metabolisms in Tom70’s regulatory function of mitochondrial biogenesis.
(A) mRNA abundance of basic mitochondrial transcription machineries quantified by RT-qPCR in TOM70 OE strain and normalized to control cells. All different yeast strains, including wild-type …
Figure 4 with 1 supplement
The Tom70-dependent regulation of mitochondrial biogenesis is involved in the cellular response to the mitochondrial import defect.
(A) Mitochondrial biogenesis in wild type cells. Metabolic signals regulate the transcriptional activity of mitochondrial proteins, which are synthesized in the cytosol and translocated through the …
Figure 4—figure supplement 1
Model of Tom70’s role in the cellular response to the mitochondrial import defect.
(A) Schematic of mitochondrial import steps involve Hsp70 and Tom70. In yeast, Hsp70 binds and maintains nascent mitochondrial proteins in unfolded states before transferring them to Tom70 for …
Figure 5 with 1 supplement
Age-related reduction of Tom70 is associated with mitochondrial dysfunctions.
(A) Representative images of Tom70-GFP and mito-RFP in young and aged cells. Both young and old cells were from YPD culture and stained with calcofluor white to visualize the bud scars. (B) …
Figure 5—figure supplement 1
Overexpressing Tom70 from a different promoter can rescue age-associated mitochondrial defects.
(A, B) Quantification of mitochondrial proteins abundance (A) and membrane potential (B) in different cells. The age-associated loss of mitochondrial membrane potential and protein expression can be …
Figure 6 with 1 supplement
Loss of Tom70 accelerates mitochondrial aging.
(A, B) Representative images (A) and quantifications (B) of Atp25-GFP in young and aged cells of different strains. Both young and old cells were from YPD culture and stained with calcofluor white …
Figure 6—figure supplement 1
Additional data for the accelerated loss of mitochondrial proteins in ∆tom70 cells during aging.
Quantifications of different mitochondrial proteins in young and aged cells of different strains. Cells with advanced reproductive age were purified from YPD culture and stained with calcofluor …
Figure 7 with 1 supplement
Reduced biogenesis and enhanced degradation underlying the age-dependent reduction of mitochondrial Tom70.
(A) Mean and s.e.m. of Tom70-GFP signal in young and old cells from strains overexpressing different TFs. Sample sizes are given in Supplementary file 4. Both young and old cells were from …
Figure 7—figure supplement 1
Additional data for the mechanisms that control the expression of Tom70 in young and aged cells.
(A) Expressing TOM70-GFP from pZ3EV can prevent the age-associated reduction of Tom70 protein. Both young and old cells were from YPD medium with 10 nM β-estradiol. (B) Tom70 mRNA quantified from …
Author response image 1
The role of mtDNA metabolisms in Tom70’s regulatory function of mitochondrial biogenesis.
(A) mRNA abundance of basic mitochondrial transcription machineries quantified by qPCR in TOM70 OE strain and normalized to control cells. All different yeast strains, including wild type control, …
Author response image 2
Quantification of different mitochondrial proteins in wild type control strain and TOM70 OE strains with/without NAC (20mM) treatment.
All different yeast strains were cultured in the same growth medium. ROS scavenger NAC partially inhibits the induction of some mitochondrial proteins (highlighted as yellow bars) in TOM70 OE cells. …
Author response image 3
Schematic of mitochondrial import steps involve Hsp70 and Tom70.
In yeast, Hsp70 binds and maintains nascent mitochondrial proteins in unfolded states before transferring them to Tom70 for import (Young et al., 2003). Tim23, the key subunits of the inner membrane …
Author response image 4
Representative WB of WT and TOM70 OE strains.
Additional files
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Supplementary file 1
Experimental models, organisms and strains.
- https://cdn.elifesciences.org/articles/75658/elife-75658-supp1-v2.xlsx
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Supplementary file 2
Oligo DNA.
- https://cdn.elifesciences.org/articles/75658/elife-75658-supp2-v2.xlsx
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Supplementary file 3
Recombinant DNA.
- https://cdn.elifesciences.org/articles/75658/elife-75658-supp3-v2.xlsx
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Supplementary file 4
Source data for sample sizes related to several figures.
- https://cdn.elifesciences.org/articles/75658/elife-75658-supp4-v2.xlsx
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Supplementary file 5
Oligo DNA for fly genes tested.
- https://cdn.elifesciences.org/articles/75658/elife-75658-supp5-v2.xlsx
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Transparent reporting form
- https://cdn.elifesciences.org/articles/75658/elife-75658-transrepform1-v2.docx
