Felix Boos

All Publications

• The unfolded protein response of the endoplasmic reticulum supports mitochondrial biogenesis by buffering non-imported proteins. Molecular biology of the cell Knöringer, K., Groh, C., Krämer, L., Stein, K. C., Hansen, K. G., Zimmermann, J., Morgan, B., Herrmann, J. M., Frydman, J., Boos, F. 2023: mbcE23050205

  Abstract

  Almost all mitochondrial proteins are synthesized in the cytosol and subsequently targeted to mitochondria. The accumulation of non-imported precursor proteins occurring upon mitochondrial dysfunction can challenge cellular protein homeostasis. Here we show that blocking protein translocation into mitochondria results in the accumulation of mitochondrial membrane proteins at the endoplasmic reticulum, thereby triggering the unfolded protein response (UPRER). Moreover, we find that mitochondrial membrane proteins are also routed to the ER under physiological conditions. The level of ER-resident mitochondrial precursors is enhanced by import defects as well as metabolic stimuli that increase the expression of mitochondrial proteins. Under such conditions, the UPRER is crucial to maintain protein homeostasis and cellular fitness. We propose the ER serves as a physiological buffer zone for those mitochondrial precursors that cannot be immediately imported into mitochondria while engaging the UPRER to adjust the ER proteostasis capacity to the extent of precursor accumulation.

  View details for DOI 10.1091/mbc.E23-05-0205

  View details for PubMedID 37379206

• Mitochondrial dysfunction rapidly modulates the abundance and thermal stability of cellular proteins. Life science alliance Groh, C., Haberkant, P., Stein, F., Filbeck, S., Pfeffer, S., Savitski, M. M., Boos, F., Herrmann, J. M. 2023; 6 (6)

  Abstract

  Cellular functionality relies on a well-balanced, but highly dynamic proteome. Dysfunction of mitochondrial protein import leads to the cytosolic accumulation of mitochondrial precursor proteins which compromise cellular proteostasis and trigger a mitoprotein-induced stress response. To dissect the effects of mitochondrial dysfunction on the cellular proteome as a whole, we developed pre-post thermal proteome profiling. This multiplexed time-resolved proteome-wide thermal stability profiling approach with isobaric peptide tags in combination with a pulsed SILAC labelling elucidated dynamic proteostasis changes in several dimensions: In addition to adaptations in protein abundance, we observed rapid modulations of the thermal stability of individual cellular proteins. Different functional groups of proteins showed characteristic response patterns and reacted with group-specific kinetics, allowing the identification of functional modules that are relevant for mitoprotein-induced stress. Thus, our new pre-post thermal proteome profiling approach uncovered a complex response network that orchestrates proteome homeostasis in eukaryotic cells by time-controlled adaptations of the abundance and the conformation of proteins.

  View details for DOI 10.26508/lsa.202201805

  View details for PubMedID 36941057

• The African Turquoise Killifish: A Scalable Vertebrate Model for Aging and Other Complex Phenotypes. Cold Spring Harbor protocols Boos, F., Chen, J., Brunet, A. 2023

  Abstract

  The African turquoise killifish Nothobranchius furzeri is currently the shortest-lived vertebrate that can be bred in captivity. Because of its short life span of only 4-6 months, rapid generation time, high fecundity, and low cost of maintenance, the African turquoise killifish has emerged as an attractive model organism that combines the scalability of invertebrate models with the unique features of vertebrate organisms. A growing community of researchers is using the African turquoise killifish for studies in diverse fields, including aging, organ regeneration, development, "suspended animation," evolution, neuroscience, and disease. A wide range of techniques is now available for killifish research, from genetic manipulations and genomic tools to specialized assays for studying life span, organ biology, response to injury, etc. This protocol collection provides detailed descriptions of the methods that are generally applicable to all killifish laboratories and those that are limited to specific disciplines. Here, we give an overview of the features that render the African turquoise killifish a unique fast-track vertebrate model organism.

  View details for DOI 10.1101/pdb.over107737

  View details for PubMedID 37100468

• MitoStores: chaperone-controlled protein granules store mitochondrial precursors in the cytosol EMBO JOURNAL Kraemer, L., Dalheimer, N., Raeschle, M., Storchova, Z., Pielage, J., Boos, F., Herrmann, J. M. 2023: e112309

  Abstract

  Hundreds of nucleus-encoded mitochondrial precursor proteins are synthesized in the cytosol and imported into mitochondria in a post-translational manner. However, the early processes associated with mitochondrial protein targeting remain poorly understood. Here, we show that in Saccharomyces cerevisiae, the cytosol has the capacity to transiently store mitochondrial matrix-destined precursors in dedicated deposits that we termed MitoStores. Competitive inhibition of mitochondrial protein import via clogging of import sites greatly enhances the formation of MitoStores, but they also form during physiological cell growth on nonfermentable carbon sources. MitoStores are enriched for a specific subset of nucleus-encoded mitochondrial proteins, in particular those containing N-terminal mitochondrial targeting sequences. Our results suggest that MitoStore formation suppresses the toxic potential of aberrantly accumulating mitochondrial precursor proteins and is controlled by the heat shock proteins Hsp42 and Hsp104. Thus, the cytosolic protein quality control system plays an active role during the early stages of mitochondrial protein targeting through the coordinated and localized sequestration of mitochondrial precursor proteins.

  View details for DOI 10.15252/embj.2022112309

  View details for Web of Science ID 000921294500001

  View details for PubMedID 36704946

• Widespread use of unconventional targeting signals in mitochondrial ribosome proteins EMBO JOURNAL Bykov, Y. S., Flohr, T., Boos, F., Zung, N., Herrmann, J. M., Schuldiner, M. 2021: e109519

  Abstract

  Mitochondrial ribosomes are complex molecular machines indispensable for respiration. Their assembly involves the import of several dozens of mitochondrial ribosomal proteins (MRPs), encoded in the nuclear genome, into the mitochondrial matrix. Proteomic and structural data as well as computational predictions indicate that up to 25% of yeast MRPs do not have a conventional N-terminal mitochondrial targeting signal (MTS). We experimentally characterized a set of 15 yeast MRPs in vivo and found that five use internal MTSs. Further analysis of a conserved model MRP, Mrp17/bS6m, revealed the identity of the internal targeting signal. Similar to conventional MTS-containing proteins, the internal sequence mediates binding to TOM complexes. The entire sequence of Mrp17 contains positive charges mediating translocation. The fact that these sequence properties could not be reliably predicted by standard methods shows that mitochondrial protein targeting is more versatile than expected. We hypothesize that structural constraints imposed by ribosome assembly interfaces may have disfavored N-terminal presequences and driven the evolution of internal targeting signals in MRPs.

  View details for DOI 10.15252/embj.2021109519

  View details for Web of Science ID 000719290500001

  View details for PubMedID 34786732

• Increased levels of mitochondrial import factor Mia40 prevent the aggregation of polyQ proteins in the cytosol. The EMBO journal Schlagowski, A. M., Knöringer, K., Morlot, S., Sánchez Vicente, A., Flohr, T., Krämer, L., Boos, F., Khalid, N., Ahmed, S., Schramm, J., Murschall, L. M., Haberkant, P., Stein, F., Riemer, J., Westermann, B., Braun, R. J., Winklhofer, K. F., Charvin, G., Herrmann, J. M. 2021: e107913

  Abstract

  The formation of protein aggregates is a hallmark of neurodegenerative diseases. Observations on patient samples and model systems demonstrated links between aggregate formation and declining mitochondrial functionality, but causalities remain unclear. We used Saccharomyces cerevisiae to analyze how mitochondrial processes regulate the behavior of aggregation-prone polyQ protein derived from human huntingtin. Expression of Q97-GFP rapidly led to insoluble cytosolic aggregates and cell death. Although aggregation impaired mitochondrial respiration only slightly, it considerably interfered with the import of mitochondrial precursor proteins. Mutants in the import component Mia40 were hypersensitive to Q97-GFP, whereas Mia40 overexpression strongly suppressed the formation of toxic Q97-GFP aggregates both in yeast and in human cells. Based on these observations, we propose that the post-translational import of mitochondrial precursor proteins into mitochondria competes with aggregation-prone cytosolic proteins for chaperones and proteasome capacity. Mia40 regulates this competition as it has a rate-limiting role in mitochondrial protein import. Therefore, Mia40 is a dynamic regulator in mitochondrial biogenesis that can be exploited to stabilize cytosolic proteostasis.

  View details for DOI 10.15252/embj.2021107913

  View details for PubMedID 34191328

• The chaperone-binding activity of the mitochondrial surface receptor Tom70 protects the cytosol against mitoprotein-induced stress. Cell reports Backes, S., Bykov, Y. S., Flohr, T., Raschle, M., Zhou, J., Lenhard, S., Kramer, L., Muhlhaus, T., Bibi, C., Jann, C., Smith, J. D., Steinmetz, L. M., Rapaport, D., Storchova, Z., Schuldiner, M., Boos, F., Herrmann, J. M. 2021; 35 (1): 108936

  Abstract

  Most mitochondrial proteins are synthesized as precursors in the cytosol and post-translationally transported into mitochondria. The mitochondrial surface protein Tom70 acts at the interface of the cytosol and mitochondria. Invitro import experiments identified Tom70 as targeting receptor, particularly for hydrophobic carriers. Using invivo methods and high-content screens, we revisit the question of Tom70 function and considerably expand the set of Tom70-dependent mitochondrial proteins. We demonstrate that the crucial activity of Tom70 is its ability to recruit cytosolic chaperones to the outer membrane. Indeed, tethering an unrelated chaperone-binding domain onto the mitochondrial surface complements most of the defects caused by Tom70 deletion. Tom70-mediated chaperone recruitment reduces the proteotoxicity of mitochondrial precursor proteins, particularly of hydrophobic inner membrane proteins. Thus, our work suggests that the predominant function of Tom70 is to tether cytosolic chaperones to the outer mitochondrial membrane, rather than to serve as a mitochondrion-specifying targeting receptor.

  View details for DOI 10.1016/j.celrep.2021.108936

  View details for PubMedID 33826901

• More than just a ticket canceller: the mitochondrial processing peptidase tailors complex precursor proteins at internal cleavage sites MOLECULAR BIOLOGY OF THE CELL Friedl, J., Knopp, M. R., Groh, C., Paz, E., Gould, S. B., Herrmann, J. M., Boos, F. 2020; 31 (24): 2657-2668

  Abstract

  Most mitochondrial proteins are synthesized as precursors that carry N-terminal presequences. After they are imported into mitochondria, these targeting signals are cleaved off by the mitochondrial processing peptidase (MPP). Using the mitochondrial tandem protein Arg5,6 as a model substrate, we demonstrate that MPP has an additional role in preprotein maturation, beyond the removal of presequences. Arg5,6 is synthesized as a polyprotein precursor that is imported into mitochondria and subsequently separated into two distinct enzymes. This internal processing is performed by MPP, which cleaves the Arg5,6 precursor at its N-terminus and at an internal site. The peculiar organization of Arg5,6 is conserved across fungi and reflects the polycistronic arginine operon in prokaryotes. MPP cleavage sites are also present in other mitochondrial fusion proteins from fungi, plants, and animals. Hence, besides its role as a "ticket canceller" for removal of presequences, MPP exhibits a second conserved activity as an internal processing peptidase for complex mitochondrial precursor proteins.

  View details for DOI 10.1091/mbc.E20-08-0524

  View details for Web of Science ID 000590617600005

  View details for PubMedID 32997570

• The intermembrane space protein Mix23 is a novel stress-induced mitochondrial import factor. The Journal of biological chemistry Zöller, E., Laborenz, J., Krämer, L., Boos, F., Räschle, M., Alexander, R. T., Herrmann, J. M. 2020; 295 (43): 14686-14697

  Abstract

  The biogenesis of mitochondria requires the import of hundreds of precursor proteins. These proteins are transported post-translationally with the help of chaperones, meaning that the overproduction of mitochondrial proteins or the limited availability of chaperones can lead to the accumulation of cytosolic precursor proteins. This imposes a severe challenge to cytosolic proteostasis and triggers a specific transcription program called the mitoprotein-induced stress response, which activates the proteasome system. This coincides with the repression of mitochondrial proteins, including many proteins of the intermembrane space. In contrast, herein we report that the so-far-uncharacterized intermembrane space protein Mix23 is considerably up-regulated when mitochondrial import is perturbed. Mix23 is evolutionarily conserved and a homolog of the human protein CCDC58. We found that, like the subunits of the proteasome, Mix23 is under control of the transcription factor Rpn4. It is imported into mitochondria by the mitochondrial disulfide relay. Mix23 is critical for the efficient import of proteins into the mitochondrial matrix, particularly if the function of the translocase of the inner membrane 23 is compromised such as in temperature-sensitive mutants of Tim17. Our observations identify Mix23 as a novel regulator or stabilizer of the mitochondrial protein import machinery that is specifically up-regulated upon mitoprotein-induced stress conditions.

  View details for DOI 10.1074/jbc.RA120.014247

  View details for PubMedID 32826315

  View details for PubMedCentralID PMC7586232

• How the Mitoprotein-Induced Stress Response Safeguards the Cytosol: A Unified View. Trends in cell biology Boos, F., Labbadia, J., Herrmann, J. M. 2020; 30 (3): 241-254

  Abstract

  Mitochondrial and cytosolic proteostasis are of central relevance for cellular stress resistance and organismal health. Recently, a number of individual cellular programs were described that counter the fatal consequences of mitochondrial dysfunction. These programs remove arrested import intermediates from mitochondrial protein translocases, stabilize protein homeostasis within mitochondria, and, in particular, increase the levels and activity of chaperones and the proteasome system in the cytosol. Here, we describe the different responses to mitochondrial perturbation and propose to unify the seemingly distinct mitochondrial-cytosolic quality control mechanisms into a single network, the mitoprotein-induced stress response. This holistic view places mitochondrial biogenesis at a central position of the cellular proteostasis network, emphasizing the importance of mitochondrial protein import processes for development, reproduction, and ageing.

  View details for DOI 10.1016/j.tcb.2019.12.003

  View details for PubMedID 31964548

• The NADH Dehydrogenase Nde1 Executes Cell Death after Integrating Signals from Metabolism and Proteostasis on the Mitochondrial Surface MOLECULAR CELL Saladi, S., Boos, F., Poglitsch, M., Meyer, H., Sommer, F., Muehlhaus, T., Schroda, M., Schuldiner, M., Madeo, F., Herrmann, J. M. 2020; 77 (1): 189-+

  Abstract

  The proteolytic turnover of mitochondrial proteins is poorly understood. Here, we used a combination of dynamic isotope labeling and mass spectrometry to gain a global overview of mitochondrial protein turnover in yeast cells. Intriguingly, we found an exceptionally high turnover of the NADH dehydrogenase, Nde1. This homolog of the mammalian apoptosis inducing factor, AIF, forms two distinct topomers in mitochondria, one residing in the intermembrane space while the other spans the outer membrane and is exposed to the cytosol. The surface-exposed topomer triggers cell death in response to pro-apoptotic stimuli. The surface-exposed topomer is degraded by the cytosolic proteasome/Cdc48 system and the mitochondrial protease Yme1; however, it is strongly enriched in respiratory-deficient cells. Our data suggest that in addition to their role in electron transfer, mitochondrial NADH dehydrogenases such as Nde1 or AIF integrate signals from energy metabolism and cytosolic proteostasis to eliminate compromised cells from growing populations.

  View details for DOI 10.1016/j.molcel.2019.09.027

  View details for Web of Science ID 000505192900016

  View details for PubMedID 31668496

• Multiple mitochondrial thioesterases have distinct tissue and substrate specificity and CoA regulation, suggesting unique functional roles. The Journal of biological chemistry Bekeova, C., Anderson-Pullinger, L., Boye, K., Boos, F., Sharpadskaya, Y., Herrmann, J. M., Seifert, E. L. 2019; 294 (50): 19034-19047

  Abstract

  Acyl-CoA thioesterases (Acots) hydrolyze fatty acyl-CoA esters. Acots in the mitochondrial matrix are poised to mitigate β-oxidation overload and maintain CoA availability. Several Acots associate with mitochondria, but whether they all localize to the matrix, are redundant, or have different roles is unresolved. Here, we compared the suborganellar localization, activity, expression, and regulation among mitochondrial Acots (Acot2, -7, -9, and -13) in mitochondria from multiple mouse tissues and from a model of Acot2 depletion. Acot7, -9, and -13 localized to the matrix, joining Acot2 that was previously shown to localize there. Mitochondria from heart, skeletal muscle, brown adipose tissue, and kidney robustly expressed Acot2, -9, and -13; Acot9 levels were substantially higher in brown adipose tissue and kidney mitochondria, as was activity for C4:0-CoA, a unique Acot9 substrate. In all tissues, Acot2 accounted for about half of the thioesterase activity for C14:0-CoA and C16:0-CoA. In contrast, liver mitochondria from fed and fasted mice expressed little Acot activity, which was confined to long-chain CoAs and due mainly to Acot7 and Acot13 activities. Matrix Acots occupied different functional niches, based on substrate specificity (Acot9 versus Acot2 and -13) and strong CoA inhibition (Acot7, -9, and -13, but not Acot2). Interpreted in the context of β-oxidation, CoA inhibition would prevent Acot-mediated suppression of β-oxidation, while providing a release valve when CoA is limiting. In contrast, CoA-insensitive Acot2 could provide a constitutive siphon for long-chain fatty acyl-CoAs. These results reveal how the family of matrix Acots can mitigate β-oxidation overload and prevent CoA limitation.

  View details for DOI 10.1074/jbc.RA119.010901

  View details for PubMedID 31676684

  View details for PubMedCentralID PMC6916504

• Mitochondrial protein translocation-associated degradation NATURE Martensson, C. U., Priesnitz, C., Song, J., Ellenrieder, L., Doan, K., Boos, F., Floerchinger, A., Zufall, N., Oeljeklaus, S., Warscheid, B., Becker, T. 2019; 569 (7758): 679-+

  Abstract

  Mitochondrial biogenesis and functions depend on the import of precursor proteins via the 'translocase of the outer membrane' (TOM complex). Defects in protein import lead to an accumulation of mitochondrial precursor proteins that induces a range of cellular stress responses. However, constitutive quality-control mechanisms that clear trapped precursor proteins from the TOM channel under non-stress conditions have remained unknown. Here we report that in Saccharomyces cerevisiae Ubx2, which functions in endoplasmic reticulum-associated degradation, is crucial for this quality-control process. A pool of Ubx2 binds to the TOM complex to recruit the AAA ATPase Cdc48 for removal of arrested precursor proteins from the TOM channel. This mitochondrial protein translocation-associated degradation (mitoTAD) pathway continuously monitors the TOM complex under non-stress conditions to prevent clogging of the TOM channel with precursor proteins. The mitoTAD pathway ensures that mitochondria maintain their full protein-import capacity, and protects cells against proteotoxic stress induced by impaired transport of proteins into mitochondria.

  View details for DOI 10.1038/s41586-019-1227-y

  View details for Web of Science ID 000470144100036

  View details for PubMedID 31118508

• Mitochondrial protein-induced stress triggers a global adaptive transcriptional programme NATURE CELL BIOLOGY Boos, F., Kraemer, L., Groh, C., Jung, F., Haberkant, P., Stein, F., Wollweber, F., Gackstatter, A., Zoeller, E., van der Laan, M., Savitski, M. M., Benes, V., Herrmann, J. M. 2019; 21 (4): 442-+

  Abstract

  The cytosolic accumulation of mitochondrial precursors is hazardous to cellular fitness and is associated with a number of diseases. However, it is not observed under physiological conditions. Individual mechanisms that allow cells to avoid cytosolic accumulation of mitochondrial precursors have recently been discovered, but their interplay and regulation remain elusive. Here, we show that cells rapidly launch a global transcriptional programme to restore cellular proteostasis after induction of a 'clogger' protein that reduces the number of available mitochondrial import sites. Cells upregulate the protein folding and proteolytic systems in the cytosol and downregulate both the cytosolic translation machinery and many mitochondrial metabolic enzymes, presumably to relieve the workload of the overstrained mitochondrial import system. We show that this transcriptional remodelling is a combination of a 'wideband' core response regulated by the transcription factors Hsf1 and Rpn4 and a unique mitoprotein-induced downregulation of the oxidative phosphorylation components, mediated by an inactivation of the HAP complex.

  View details for DOI 10.1038/s41556-019-0294-5

  View details for Web of Science ID 000462859500007

  View details for PubMedID 30886345

• Detection of Internal Matrix Targeting Signal-like Sequences (iMTS-Ls) in Mitochondrial BIO-PROTOCOL Boos, F., Muehlhaus, T., Herrmann, J. M. 2018; 8 (17)

  View details for DOI 10.21769/BioProtoc.2474

  View details for Web of Science ID 000458021000003

• Genome-wide SWAp-Tag yeast libraries for proteome exploration NATURE METHODS Weill, U., Yofe, I., Sass, E., Stynen, B., Davidi, D., Natarajan, J., Ben-Menachem, R., Avihou, Z., Goldman, O., Harpaz, N., Chuartzman, S., Kniazev, K., Knoblach, B., Laborenz, J., Boos, F., Kowarzyk, J., Ben-Dor, S., Zalckvar, E., Herrmann, J. M., Rachubinski, R. A., Pines, O., Rapaport, D., Michnick, S. W., Levy, E. D., Schuldiner, M. 2018; 15 (8): 617-+

  Abstract

  Yeast libraries revolutionized the systematic study of cell biology. To extensively increase the number of such libraries, we used our previously devised SWAp-Tag (SWAT) approach to construct a genome-wide library of ~5,500 strains carrying the SWAT NOP1promoter-GFP module at the N terminus of proteins. In addition, we created six diverse libraries that restored the native regulation, created an overexpression library with a Cherry tag, or enabled protein complementation assays from two fragments of an enzyme or fluorophore. We developed methods utilizing these SWAT collections to systematically characterize the yeast proteome for protein abundance, localization, topology, and interactions.

  View details for DOI 10.1038/s41592-018-0044-9

  View details for Web of Science ID 000440334000023

  View details for PubMedID 29988094

  View details for PubMedCentralID PMC6076999

• Accessory signals in protein translocation AGING-US Hansen, K. G., Boos, F., Herrmann, J. M. 2018; 10 (4): 530-531

  View details for DOI 10.18632/aging.101435

  View details for Web of Science ID 000433595400008

  View details for PubMedID 29706613

  View details for PubMedCentralID PMC5940120

• Tom70 enhances mitochondrial preprotein import efficiency by binding to internal targeting sequences JOURNAL OF CELL BIOLOGY Backes, S., Hess, S., Boos, F., Woellhaf, M. W., Goedel, S., Jung, M., Muehlhaus, T., Herrmann, J. M. 2018; 217 (4): 1369-1382

  Abstract

  The biogenesis of mitochondria depends on the import of hundreds of preproteins. N-terminal matrix-targeting signals (MTSs) direct preproteins to the surface receptors Tom20, Tom22, and Tom70. In this study, we show that many preproteins contain additional internal MTS-like signals (iMTS-Ls) in their mature region that share the characteristic properties of presequences. These features allow the in silico prediction of iMTS-Ls. Using Atp1 as model substrate, we show that iMTS-Ls mediate the binding to Tom70 and have the potential to target the protein to mitochondria if they are presented at its N terminus. The import of preproteins with high iMTS-L content is significantly impaired in the absence of Tom70, whereas preproteins with low iMTS-L scores are less dependent on Tom70. We propose a stepping stone model according to which the Tom70-mediated interaction with internal binding sites improves the import competence of preproteins and increases the efficiency of their translocation into the mitochondrial matrix.

  View details for DOI 10.1083/jcb.201708044

  View details for Web of Science ID 000428997800019

  View details for PubMedID 29382700

  View details for PubMedCentralID PMC5881500

• Methionine on the rise: how mitochondria changed their codon usage. The EMBO journal Boos, F., Wollin, M., Herrmann, J. M. 2016; 35 (19): 2066-2067

  View details for DOI 10.15252/embj.201695385

  View details for PubMedID 27578810

  View details for PubMedCentralID PMC5048349