1. Volume 42, Issue 3, Pages 390-400 (May 2011)
Absence of Mitochondrial Translation Control Proteins Extends Life Span by Activating Sirtuin-Dependent Silencing 
Antonio Caballero, Ana Ugidos, Beidong Liu, David Öling, Kristian Kvint, Xinxin Hao, Cora Mignat, Laurence Nachin, Mikael Molin, Thomas Nyström 
Molecular Cell 
Volume 42, Issue 3, Pages (May 2011)
DOI: /j.molcel
Copyright © 2011 Elsevier Inc. Terms and Conditions

2. Figure 1
A Deletion of SOV1 Extends Life Span without Affecting Levels of ROS or Protein Oxidation
(A) Replicative life span (RLS) of wild-type in black (mean RLS = 24; standard error [SE] ± 0.89 n = 154) and sov1Δ in red (mean RLS = 32; SE ± 1.8 n = 97). The plot is the average of three different experiments. Levels of superoxides (B) and peroxides (C) in exponential growing wild-type (Wt) and sov1Δ cells, measured as the intensity of Dihydroethidium (DHE) and Dihydrorhodamine (DHR) signal, respectively, by flow cytometry. Levels of superoxides and peroxides were calculated using the Flowjo software and plotted as mean values ± SD. (D) Pattern of carbonylated proteins in Wt and sov1Δ analyzed by anti-DNPH western blotting of 2D-PAGE separated protein derivatized with DNPH. The inset show total carbonyl levels in the Wt (W) and sov1Δ (S) mutant quantified using the Image Gauge software (FUJI) and normalized to total protein. See also Figure S1.
Molecular Cell  , DOI: ( /j.molcel )
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3. Figure 2
RLS Extension Is Not a General Effect of Respiration Deficiency, mtDNA Loss, or Mitochondrial Protein Assembly Defects
(A) Replicative life span (RLS) of respiratory-deficient mutants, lacking COX6 (blue, mean RLS = 23; SE ± 1.34 n = 60), COX7 (green, mean RLS = 18; SE ± 1.17 n = 49), and SHY1 (orange, mean RLS = 26; SE ± 1.46 n = 54) compared to the Wt (black, mean RLS = 24; SE ± 0.89 n = 154).
(B) RLS of sov1Δmip1Δ cells (red, mean RLS = 18; SE ± 0.62 n = 124) and mip1Δ cells (blue, mean RLS = 15; SE ± 0.64 n = 125).
(C) DAPI staining of nuclear and mtDNA in Wt and mip1Δ cells as indicated. See also Figure S2.
Molecular Cell  , DOI: ( /j.molcel )
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4. Figure 3
SOV1 Interacts Genetically with Genes Involved in Chromatin Remodeling
(A) Functional categories enriched among genes genetically interacting with SOV1.
(B) A network of complexes showing synthetic sick/lethal interactions with SOV1. Genes showing genetic interactions with SOV1 (nodes) were grouped in modules based on their known physical interactions (blue edges). Edges between SOV1 and complexes are the genetic interactions found in this study and colors show the strength of the interactions (gray, SS, synthetic sick; red, SSS, severe synthetic sick; black, SL, synthetic lethal).
Molecular Cell  , DOI: ( /j.molcel )
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5. Figure 4 SOV1 Affects Sir2-Dependent Silencing and Life Span Control
(A) rDNA silencing measured as expression of the URA3 gene located in the rDNA loci in Wt, sov1Δ, sir2Δ, and the double mutant sir2Δsov1Δ in drop test assay in YNB/YNB-Ura media.
(B) Silencing at mating type loci was assayed in Wt and sov1Δ in a HMR::TRP1 mutation background strain.
(C) RLS of the sir2Δsov1Δ mutant (blue line, mean RLS = 13; SE ± 0.56 n = 79), and the sir2Δ (orange, mean RLS = 13; SE ± 0.56 n = 56) mutant.
(D) RLS of a triple sov1Δsir2Δfob1Δ mutant (yellow line, mean RLS = 21; SE ± 0.92 n = 124) and a sir2Δfob1Δ (blue line, mean RLS = 19; SE ± 1.43 n = 80) mutant.
(E) RLS of a double sov1Δfob1Δ (orange line, mean RLS = 35; SE ± 1.59 n = 119) mutant and the single fob1Δ (blue line, mean RLS = 35; SE ± 2.16 n = 56) mutant. See also Figure S3.
Molecular Cell  , DOI: ( /j.molcel )
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6. Figure 5
Increased Nuclear Silencing and Life Span Extension of sov1Δ Require MSN2/MSN4 and PNC1
(A) Pictures of colonies formed after random sporulation in selective minimal medium of the Wt, sov1Δ, ira2Δ, sov1Δira2Δ, ras2Δ, and sov1Δras2Δ mutant strains obtained by SGA procedure.
(B) cAMP levels in sov1Δ, ras2Δ, and sov1Δras2Δ mutants compared to the Wt. Data are represented as mean ± SD.
(C) Percentage of isolated mother cells with Msn2-GFP protein localized in the nucleus (age was determined by calcofluor, and Msn2-GFP localization profiles were determined for Wt and sov1Δ mother cells older than six generations). Data are represented as mean ± SD.
(D) Expression of HSP12 as measured by qPCR in Wt, sov1Δ, msn2/4Δ, and sov1msn2/4Δ strains. Levels were normalized to values of the ACT1 reference gene, and the Wt value was set to 1. Data are represented as mean ± SD.
(E) RLS assays of the wild-type (black line, mean RLS = 24; SE ± 0.89 n = 154), the msn2/4Δ (orange line, mean RLS = 23; SE ± 1.04 n = 109), and the sov1Δmsn2/4Δ mutant strains (blue line, mean RLS = 19; SE ± 0.86 n = 115).
(F) Silencing at the rDNA locus as measured by the URA3 gene located in the rDNA region of the Wt, sov1Δ, pnc1Δ, and pnc1Δsov1Δ strains.
(G) RLS of a pnc1Δsov1Δ mutant (orange, mean RLS = 16; SE ± 0.93 n = 54), and pnc1Δ mutant (blue, mean RLS = 21; SE ± 0.94 n = 119) compared to Wt (black line, mean RLS = 24; SE ± 0.89 n = 154).
(H and I) (H) Silencing at the rDNA locus measured by the URA3 reporter in Wt, sov1Δ, pde2Δ, and pde2Δsov1Δ strains, and (I) in ras2Δ and ras2Δsov1Δ compared to Wt and sov1Δ strains.
(J and K) (J) RLS of a ras2Δsov1Δ mutant (red line, mean RLS = 25; SE ± 0.83 n = 104), ras2Δ mutant (black line, mean RLS = 22.5; SE ± 0.93 n = 104) and (K) of pde2Δsov1Δ (green line, mean RLS = 7; SE ± 0.49 n = 40), pde2Δ (red line, mean RLS = 23.5; SE ± 1.96 n = 40) compared to Wt (black line, mean RLS = 29; SE ± 1.87 n = 40). See also Figure S4.
Molecular Cell  , DOI: ( /j.molcel )
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7. Figure 6
Mitochondrial Translation Control Module Proteins Control Life Span in a SIR2-Dependent Manner
(A) Sov1p belongs to the MTC cluster involved in the regulation of translation of mitochondrial encoded genes. The MTC module includes proteins required for translation of mitochondrial encoded COX subunits (violet), COB (pink), OLI1 (green) translation, and also both subunits of the degradosome complex (black).
(B) Life span extension of mutants lacking different MTC proteins as indicated. Data are represented as mean ± SD.
(C) Life span of Wt (blue, mean RLS = 26; SE ± 1.2 n = 92), cbs1Δ (red, RLS = 32; SE ± 1.2, n = 105), suv3Δ (black, RLS = 33; SE ± 1.94 n = 64), and suv3Δcbs1Δ (yellow, RLS = 31; SE ± 1.8, n = 51) cells.
(D) Patterns and total levels of protein carbonylation in Wt and cbs1Δ cells.
(E) Correlation between RLS, respiration, mtDNA, and inclusion in the MTC module for the deletion mutants analyzed in this paper. Grey denotes presence and white absence. Red is increased RLS, green decreased RLS, and white no change. Pet127p was not originally included in the MTC module but is required for cytochrome B translation regulation.
(F–H) (F) Life span of cbs1Δ (blue; mean RLS = 32; SE ± 1.2, n = 105), sir2Δ (red, RLS = 12; SE ± 0.46, n = 40), and cbs1Δsir2Δ (black, RLS = 12.2; SE ± 0.46, n = 41) cells; (G) cbs1Δ (blue, RLS = 32; SE ± 1.2, n = 105), sir2Δfob1Δ (red, RLS = 19; SE ± 1.4, n = 59), and sir2Δfob1Δcbs1Δ (yellow, RLS = 18; SE ± 1.16, n = 54) cells; (H) cbs1Δ (blue, RLS = 32; SE ± 1.2, n = 105), fob1Δ (red, RLS = 42; SE ± 0.4, n = 41), and fob1Δcbs1Δ (black, RLS = 41; SE ± 1.8, n = 63) cells.
(I) Silencing at rDNA as scored by URA3 expression in the rDNA locus in the wild-type and cbs1Δ strain. See also Figure S5.
Molecular Cell  , DOI: ( /j.molcel )
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8. Figure 7
The MTC Protein Cbs2p Is Nuclear Localized and Is Required for cbs1Δ-Dependent Life Span Extension and Silencing
(A) Replicative life span of Wt (black, RLS = 26; SE ± 1.2, n = 92), a cbs2Δ mutant (orange, mean RLS = 22.8; SE ± 0.86, n = 144), a cbs1Δ mutant (red, RLS = 31.8; SE ± 1.2, n = 105), and a cbs2Δ cbs1Δ double mutant (blue, RLS = 19.8; SE ± 1.14, n = 79).
(B) Cellular localization of Cbs2p as determined by a Cbs2-GFP fusion and costaining with DAPI. Nuclear localization is marked with “N” and mitochondrial localization with “mt.” BF, bright field.
(C) Silencing at rDNA as scored by URA3 expression in the rDNA locus in the wild-type, cbs1Δ, cbs2Δ, and cbs2Δ cbs1Δ strains as indicated.
Molecular Cell  , DOI: ( /j.molcel )
Copyright © 2011 Elsevier Inc. Terms and Conditions