The mitochondrial acyl carrier protein (ACP) coordinates mitochondrial fatty acid synthesis with iron sulfur cluster biogenesis
- University of Utah School of Medicine, United States
- Harvard University School of Medicine, United States
- Howard Hughes Medical Institute, University of Utah School of Medicine, United States
Figures
Figure 1 with 3 supplements
Acp1 is a stable subunit of the ISU complex.
(A) acp1Δ heterozygous diploids were dissected with and without a plasmid expressing Acp1-HA (BY4741) and spores were grown on YPAD medium for 2 days. Sporulation of the heterozygous deletion strain …
Figure 1—figure supplement 1
Protein sequence alignment of ACP from eukaryotes.
(A) Protein sequence alignment of the ACP proteins from the indicated species (Clustal Omega). D.m. – Drosophila melanogaster, D.r. – Danio rerio, H.s. – Homo sapiens, M.m. – Mus musculus, S.c. – Sac…
Figure 1—figure supplement 2
Eukarotic mitochondrial fatty acid biosynthesis (FASII) pathway.
(A) Eukaryotic cells have maintained two distinct fatty acid synthesis pathways—the canonical cytoplasmic FASI and the mitochondrial FASII, which is homologous to the prokaryotic fatty acid …
Figure 1—figure supplement 3
Acp1 is a stable subunit of the ISU complex.
(A) Isd11-V5 and Acp1- HA were expressed from their endogenous loci and Isd11-V5 was immunoprecipitated from digitonin-solubilized mitochondria followed by the indicated immunoblots. (B) Nfs1-V5 and …
Figure 2 with 2 supplements
Acp1 is required for FeS biogenesis.
(A) Isolated mitochondria from the indicated strains were resolved by SDS-PAGE (upper panel) or solubilized in 1% digitonin and resolved by BN-PAGE (lower panel). The time course indicates the time …
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Figure 2—source data 1
Source data for Figure 2.
This file contains raw source data used to make the graphs presented in Figure 2B,C Figure 2—figure supplement 2B and C. PRISM software was used to graph all quantitative data and perform statistical analyses. p values for pairwise comparisons were determined using a Student’s t test.
- https://doi.org/10.7554/eLife.17828.008
Figure 2—figure supplement 1
ACP1 expression is required for cell proliferation.
(A) Ten-fold serial dilutions of WT, TetO7-ACP1, and TetO7-ACP1 cells expressing Acp1-HA were plated on synthetic media containing the indicated carbon source with and without 10 μg/mL doxycycline …
Figure 2—figure supplement 2
Acp1 is essential for FeS biogenesis.
(A) Isolated mitochondria from WT, TetO7-ACP1, or TetO7-ACP1 cells expressing Acp1-HA were harvested at the indicated times post addition of doxycycline were resolved by SDS-PAGE (upper panels) or …
Figure 3 with 1 supplement
ACP promotes FeS biogenesis by maintaining the stability of the ISU (Nfs1-Isd11) complex.
(A) Purified mitochondria from the indicated strains were either resolved by SDS-PAGE (lower panels) or solubilized in 1% digitonin and resolved by BN-PAGE (upper panels). Cells were grown for 18 hr …
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Figure 3—source data 1
Source data for Figure 3.
This file contains raw source data used to make the graphs presented in Figure 3—figure supplement 1C. PRISM software was used to graph all quantitative data and perform statistical analyses. p values for pairwise comparisons were determined using a Student’s t test.
- https://doi.org/10.7554/eLife.17828.012
Figure 3—figure supplement 1
ACP promotes FeS biogenesis by maintaining the stability of the ISU (Nfs1-Isd11) complex.
(A) Isolated mitochondria from WT and TetO7-ACP1 cells harvested at the indicated time points post-addition of doxycycline were resolved by SDS-PAGE (upper panel) or solubilized in 1% digitonin and …
Figure 4 with 1 supplement
Acp1 requires a 4-PP-conjugated acyl chain to fully stabilize the ISU complex.
(A) Isolated mitochondria from the indicated strains expressing the indicated genes by plasmid were either resolved by SDS-PAGE (upper panels) or solubilized in 1% digitonin and resolved by BN-PAGE …
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Figure 4—source data 1
Source data for Figure 4.
- https://doi.org/10.7554/eLife.17828.015
Figure 4—figure supplement 1
Acp1 requires a 4-PP-conjugated acyl chain to fully stabilize the ISU complex.
(A) Ten-fold serial dilutions of the indicated strains plated on synthetic medium with either glucose or glycerol. (B) Ten-fold serial dilutions of the indicated strains expressing the indicated …
Additional files
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Supplementary file 1
Yeast strains used in this study.
This table describes the name, genotype, and source of all yeast strains used in this investigation.
- https://doi.org/10.7554/eLife.17828.017
