Deletion of the natural inhibitory protein Inh1 in Ustilago maydis has no effect on the dimeric state of the F1FO-ATP synthase but increases the ATPase activity and reduces the stability

Romero-Aguilar, Lucero; Esparza-Perusquía, Mercedes; Langner, Thorsten; García-Cruz, Giovanni; Feldbrügge, Michael; Zavala, Guadalupe; Pablo, Pardo Juan; Martı́nez, Federico; Flores-Herrera, Óscar

doi:10.1016/j.bbabio.2021.148429

Cited by 4 publications

(5 citation statements)

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“…The key role of these natural F 1 F O -ATPase inhibitors has been researched by means of knockout or null IF 1 and ζ mutants, given that it is not possible to delete the essential ε subunit since its deletion leads to the detachment of F 1 from F O in bacterial F-ATP synthase because of its primary function as a central rotor subunit connecting F 1 with F O [ 31 ]. Several knockouts of mitochondrial IF 1 in fungi [ 61 ], yeast [ 62 ], nematodes [ 63 ], and mice [ 64 ] failed to show a knockout phenotype different to the wild-type strains, either because there are multiple copies of the IF 1 gene or because all of these species contain assembly or stabilizing factors genes (STFs) similar to IF 1 [ 65 , 66 , 67 , 68 ]. These IF 1 knockouts have only been made in a single IF 1 gene in most cases, and therefore, the other IF 1 gene copies or IF 1 -like stabilizing factors (STF1 or STF2) very likely complement the missing IF 1 gene in these single knockouts, thus explaining the lack of a clear different phenotype of the null mutants compared with the wild-type strains.…”

Section: Resultsmentioning

confidence: 99%

“…Our previous functional observation of the role of IF 1 in stabilizing the dimeric and oligomeric structures of the mtATP synthase [ 108 ], besides inhibiting the F 1 F O -ATPase activity, has been therefore structurally confirmed by the cryo-EM structure of the pig tetrameric F-ATP synthase [ 115 ]. Other IF 1 knockout analyses have apparently shown that IF 1 is not essential or necessary for the dimerization or oligomerization of the mitochondrial F-ATP synthase since the F-ATP synthase dimer is preserved in BN-PAGE analysis in both WT and ΔIF 1 (or ΔINH1) knockout mutants [ 61 , 63 , 64 , 117 ]. However, as discussed above, at least one IF 1 gene copy or homologous IF 1 -like gene that still remains expressed could complement or substitute the inhibiting and dimerizing function of the missing IF 1 .…”

Section: Resultsmentioning

confidence: 99%

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Evolution of the Inhibitory and Non-Inhibitory ε, ζ, and IF1 Subunits of the F1FO-ATPase as Related to the Endosymbiotic Origin of Mitochondria

et al. 2022

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The F1FO-ATP synthase nanomotor synthesizes >90% of the cellular ATP of almost all living beings by rotating in the “forward” direction, but it can also consume the same ATP pools by rotating in “reverse.” To prevent futile F1FO-ATPase activity, several different inhibitory proteins or domains in bacteria (ε and ζ subunits), mitochondria (IF1), and chloroplasts (ε and γ disulfide) emerged to block the F1FO-ATPase activity selectively. In this study, we analyze how these F1FO-ATPase inhibitory proteins have evolved. The phylogeny of the α-proteobacterial ε showed that it diverged in its C-terminal side, thus losing both the inhibitory function and the ATP-binding/sensor motif that controls this inhibition. The losses of inhibitory function and the ATP-binding site correlate with an evolutionary divergence of non-inhibitory α-proteobacterial ε and mitochondrial δ subunits from inhibitory bacterial and chloroplastidic ε subunits. Here, we confirm the lack of inhibitory function of wild-type and C-terminal truncated ε subunits of P. denitrificans. Taken together, the data show that ζ evolved to replace ε as the primary inhibitor of the F1FO-ATPase of free-living α-proteobacteria. However, the ζ inhibitory function was also partially lost in some symbiotic α-proteobacteria and totally lost in some strictly parasitic α-proteobacteria such as the Rickettsiales order. Finally, we found that ζ and IF1 likely evolved independently via convergent evolution before and after the endosymbiotic origin mitochondria, respectively. This led us to propose the ε and ζ subunits as tracer genes of the pre-endosymbiont that evolved into the actual mitochondria.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Evolution of the Inhibitory and Non-Inhibitory ε, ζ, and IF1 Subunits of the F1FO-ATPase as Related to the Endosymbiotic Origin of Mitochondria

et al. 2022

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“…The second mechanism of ATP hydrolysis inhibition is implemented with the help of natural inhibitor proteins: the ε subunit in bacteria [ 60 ] and IF1 in mitochondria [ 61 ]. This type of inhibitor also includes the ζ subunit of α-proteobacteria [ 62 ].…”

Section: Reversibility Of F 1 F O ...mentioning

confidence: 99%

“…Studies of the IF1 function in various cell lines produced conflicting results, which did not help to understand the real role of this protein in the whole organism. In some experiments, IF1 knockout increased mitochondrial ATPase activity [ 61 ]. Other IF1 knockout mutants in yeast, mice or Caenorhabditis elegans showed no difference in growth, reproduction or bioenergetics compared to wild types [ 75 ].…”

Section: Reversibility Of F 1 F O ...mentioning

confidence: 99%

F1·Fo ATP Synthase/ATPase: Contemporary View on Unidirectional Catalysis

Zharova

Grivennikova

Borisov

2023

IJMS

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F1·Fo-ATP synthases/ATPases (F1·Fo) are molecular machines that couple either ATP synthesis from ADP and phosphate or ATP hydrolysis to the consumption or production of a transmembrane electrochemical gradient of protons. Currently, in view of the spread of drug-resistant disease-causing strains, there is an increasing interest in F1·Fo as new targets for antimicrobial drugs, in particular, anti-tuberculosis drugs, and inhibitors of these membrane proteins are being considered in this capacity. However, the specific drug search is hampered by the complex mechanism of regulation of F1·Fo in bacteria, in particular, in mycobacteria: the enzyme efficiently synthesizes ATP, but is not capable of ATP hydrolysis. In this review, we consider the current state of the problem of “unidirectional” F1·Fo catalysis found in a wide range of bacterial F1·Fo and enzymes from other organisms, the understanding of which will be useful for developing a strategy for the search for new drugs that selectively disrupt the energy production of bacterial cells.

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“…In the presence of protonophores, mitochondria isolated from the yeast strain with double deletion of IF 1 genes, Δinh1Δstf1 , hydrolyze ATP much faster than the mitochondria of wild type yeast do ( Ichikawa et al, 1990 ; Venard et al, 2003 ). Moreover, deletion of IF 1 homolog increased ATP hydrolysis rate in another yeast species, Ustilago maydis ( Lucero et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Heterogeneity of Starved Yeast Cells in IF1 Levels Suggests the Role of This Protein in vivo

et al. 2022

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In mitochondria, a small protein IF1 suppresses the hydrolytic activity of ATP synthase and presumably prevents excessive ATP hydrolysis under conditions of energy deprivation. In yeast Saccharomyces cerevisiae, IF1 homologs are encoded by two paralogous genes: INH1 and STF1. INH1 expression is known to aggravate the deleterious effects of mitochondrial DNA (mtDNA) depletion. Surprisingly, no beneficial effects of INH1 and STF1 were documented for yeast so far, and the functions of INH1 and STF1 in wild type cells are unclear. Here, we put forward a hypothesis that INH1 and STF1 bring advantage during the fast start of proliferation after reentry into exponential growth from post-diauxic or stationary phases. We found that yeast cells increase the concentration of both proteins in the post-diauxic phase. Post-diauxic phase yeast cells formed two subpopulations distinct in Inh1p and Stf1p concentrations. Upon exit from the post-diauxic phase cells with high level of Inh1-GFP started growing earlier than cells devoid of Inh1-GFP. However, double deletion of INH1 and STF1 did not increase the lag period necessary for stationary phase yeast cells to start growing after reinoculation into the fresh medium. These results point to a redundancy of the mechanisms preventing uncontrolled ATP hydrolysis during energy deprivation.

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Deletion of the natural inhibitory protein Inh1 in Ustilago maydis has no effect on the dimeric state of the F1FO-ATP synthase but increases the ATPase activity and reduces the stability

Cited by 4 publications

References 47 publications

Evolution of the Inhibitory and Non-Inhibitory ε, ζ, and IF1 Subunits of the F1FO-ATPase as Related to the Endosymbiotic Origin of Mitochondria

Evolution of the Inhibitory and Non-Inhibitory ε, ζ, and IF1 Subunits of the F1FO-ATPase as Related to the Endosymbiotic Origin of Mitochondria

F1·Fo ATP Synthase/ATPase: Contemporary View on Unidirectional Catalysis

Heterogeneity of Starved Yeast Cells in IF1 Levels Suggests the Role of This Protein in vivo

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