Document S1. Two Figures and One Table

Current Biology, Volume 19
Supplemental Data
Mitochondrial Cardiolipin Involved
in Outer Membrane Protein Biogenesis:
Implications for Barth Syndrome
Natalia Gebert, Amit S. Joshi, Stephan Kutik, Thomas Becker, Matthew McKenzie,
Xue Li Guan, Ved P. Mooga, David A. Stroud, Gnanada Kulkarni, Markus R. Wenk,
Peter Rehling, Chris Meisinger, Michael T. Ryan, Nils Wiedemann,
Miriam L. Greenberg, and Nikolaus Pfanner
Figure S1. Stability and Protein Complexes of Cardiolipin Mutant Mitochondria
(A) Mitochondria were isolated from wild-type (WT), crd1Δ and taz1Δ yeast grown at 21°C. Where
indicated the mitochondria were subjected to hypotonic swelling and treatment with proteinase K (Prot. K;
50 μg/ml). The mitochondria were analyzed by SDS-PAGE and immunoblotting. Cit1, citrate synthase
(matrix protein).
(B) Isolated mitochondria were analyzed by blue native electrophoresis and immunoblotting with
antibodies directed against Tim22 or Tim10.
Figure S2. Import of Presequence-Carrying Proteins into Cardiolipin Mutant Mitochondria
Isolated yeast mitochondria were incubated with the radiolabeled precursors of (A) F1β, Su9-DHFR or (B)
Oxa1 at 24°C for the indicated time periods (min), followed by treatment with proteinase K (50 μg/ml). The
mitochondria were analyzed by SDS-PAGE and autoradiography. Quantification of imported Oxa1 was
performed by digital autoradiography and ImageQuant software (GE Healthcare). The amount of Oxa1
imported into wild-type mitochondria after 18 min was set to 100% (control) (representing ~30% of the
precursor proteins added to the import reaction). Δψ, membrane potential; m, mature; F1β, subunit β of
F1Fo-ATP synthase.
Table S1. Saccharomyces cerevisiae Strains
Strain
Genotype
YPH499 (WT)
MATa ade2-101 his3-Δ200 leu2-Δ1 ura3-52 trp1-Δ63 lys2-801
crd1Δ (YPH)
MATa ade2-101 his3-Δ200 leu2-Δ1 ura3-52 trp1-Δ63 lys2-801 crd1::kanMX4
taz1Δ (YPH)
MATa ade2-101 his3-Δ200 leu2-Δ1 ura3-52 trp1-Δ63 lys2-801 taz1::kanMX4
tam41Δ (YPH)
MATa ade2-101 his3-Δ200 leu2-Δ1 ura3-52 trp1-Δ63 lys2-801 tam41::ADE2
BY4741 (WT)
MATa his3-Δ1 leu2-Δ0 met15-Δ0 ura3-Δ0
crd1Δ (BY)
MATa his3-Δ1 leu2-Δ0 met15-Δ0 ura3-Δ0 crd1::kanMX4
taz1Δ (BY)
MATa his3-Δ1 leu2-Δ0 met15-Δ0 ura3-Δ0 taz1::kanMX4

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