Adenosine Triphosphatases; Amino Acids; Binding Sites; Conserved Sequence; Electron Transport Complex III; Humans; Membrane Proteins/genetics/physiology; Mitochondrial Proteins/genetics/physiology; Molecular Chaperones/genetics/physiology; Mutant Proteins; Point Mutation; Protein Structure, Tertiary; Saccharomyces cerevisiae Proteins/genetics/physiology
Abstract :
[en] The mitochondrial protein Bcs1p is conserved from Saccharomyces cerevisiae to humans and its C-terminal region exhibits an AAA (ATPases associated with diverse cellular activities) domain. The absence of the yeast Bcs1p leads to an assembly defect of the iron-sulfur protein (ISP) subunit within the mitochondrial respiratory complex III, whereas human point mutations located all along the protein cause various pathologies. We have performed a structure-function analysis of the yeast Bcs1p by randomly generating a collection of respiratory-deficient point mutants. We showed that most mutations are in the C-terminal region of Bcs1p and have localized them on a theoretical three-dimensional model based on the structure of several AAA proteins. The mutations can be grouped into classes according to their respiratory competence and their location on the three-dimensional model. We have further characterized five mutants, each substituting an amino acid conserved in yeast and mammalian Bcs1 proteins but not in other AAA proteins. The effects on respiratory complex assembly and Bcs1p accumulation were analyzed. Intragenic and extragenic compensatory mutations able to restore complex III assembly to the mutants affecting the AAA domain were isolated. Our results bring new insights into the role of specific residues in critical regions that are also conserved in the human Bcs1p. We show that (1) residues located at the junction between the Bcs1p-specific and the AAA domains are important for the activity and stability of the protein and (2) the residue F342 is important for interactions with other partners or substrate proteins.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Nouet, Cécile ; Université de Liège - ULiège > Département des sciences de la vie > Génomique fonctionnelle et imagerie moléculaire végétale
Truan, Gilles
Mathieu, Lise
Dujardin, Genevieve
Language :
English
Title :
Functional analysis of yeast bcs1 mutants highlights the role of Bcs1p-specific amino acids in the AAA domain.
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Bibliography
Nobrega F.G., Nobrega M.P., and Tzagoloff A. BCS1, a novel gene required for the expression of functional Rieske iron-sulfur protein in Saccharomyces cerevisiae. EMBO J. 11 (1992) 3821-3829
Petruzzella V., Tiranti V., Fernandez P., Ianna P., Carrozzo R., and Zeviani M. Identification and characterization of human cDNAs specific to BCS1, PET112, SCO1, COX15, and COX11, five genes involved in the formation and function of the mitochondrial respiratory chain. Genomics 54 (1998) 494-504
Cruciat C.M., Hell K., Folsch H., Neupert W., and Stuart R.A. Bcs1p, an AAA-family member, is a chaperone for the assembly of the cytochrome bc1 complex. EMBO J. 18 (1999) 5226-5233
de Lonlay P., Valnot I., Barrientos A., Gorbatyuk M., Tzagoloff A., Taanman J.W., et al. A mutant mitochondrial respiratory chain assembly protein causes complex III deficiency in patients with tubulopathy, encephalopathy and liver failure. Nat. Genet. 29 (2001) 57-60
Hunte C., Koepke J., Lange C., Rossmanith T., and Michel H. Structure at 2.3 Å resolution of the cytochrome bc(1) complex from the yeast Saccharomyces cerevisiae co-crystallized with an antibody Fv fragment. Structure 8 (2000) 669-684
Zara V., Conte L., and Trumpower B.L. Identification and characterization of cytochrome bc1 subcomplexes in mitochondria from yeast with single and double deletions of genes encoding cytochrome bc1 subunits. FEBS J. 274 (2007) 4526-4539
Zara V., Palmisano I., Conte L., and Trumpower B.L. Further insights into the assembly of the yeast cytochrome bc1 complex based on analysis of single and double deletion mutants lacking supernumerary subunits and cytochrome b. Eur. J. Biochem. 271 (2004) 1209-1218
Wu M., and Tzagoloff A. Identification and characterization of a new gene (CBP3) required for the expression of yeast coenzyme QH2-cytochrome c reductase. J. Biol. Chem. 264 (1989) 11122-11130
Crivellone M.D. Characterization of CBP4, a new gene essential for the expression of ubiquinol-cytochrome c reductase in Saccharomyces cerevisiae. J. Biol. Chem. 269 (1994) 21284-21292
Kronekova Z., and Rodel G. Organization of assembly factors Cbp3p and Cbp4p and their effect on bc1 complex assembly in Saccharomyces cerevisiae. Curr. Genet 47 (2005) 203-212
Visapaa I., Fellman V., Vesa J., Dasvarma A., Hutton J.L., Kumar V., et al. GRACILE syndrome, a lethal metabolic disorder with iron overload, is caused by a point mutation in BCS1L. Am. J. Hum. Genet. 71 (2002) 863-876
De Meirleir L., Seneca S., Damis E., Sepulchre B., Hoorens A., Gerlo E., et al. Clinical and diagnostic characteristics of complex III deficiency due to mutations in the BCS1L gene. Am. J. Med. Genet. A 121 (2003) 126-131
Hinson J.T., Fantin V.R., Schonberger J., Breivik N., Siem G., McDonough B., et al. Missense mutations in the BCS1L gene as a cause of the Bjornstad syndrome. N. Engl. J. Med. 356 (2007) 809-819
Fernandez-Vizarra E., Bugiani M., Goffrini P., Carrara F., Farina L., Procopio E., et al. Impaired complex III assembly associated with BCS1L gene mutations in isolated mitochondrial encephalopathy. Hum. Mol. Genet. 16 (2007) 1241-1252
Tamai S., Iida H., Yokota S., Sayano T., Kiguchiya S., Ishihara N., et al. Characterization of the mitochondrial protein LETM1, which maintains the mitochondrial tubular shapes and interacts with the AAA-ATPase BCS1L. J. Cell Sci. 121 (2008) 2588-2600
Kotarsky H., Tabasum I., Mannisto S., Heikinheimo M., Hansson S., and Fellman V. BCS1L is expressed in critical regions for neural development during ontogenesis in mice. Gene Expr. Patterns 7 (2007) 266-273
Vandenabeele S., Van Der Kelen K., Dat J., Gadjev I., Boonefaes T., Morsa S., et al. A comprehensive analysis of hydrogen peroxide-induced gene expression in tobacco. Proc. Natl Acad. Sci. USA 100 (2003) 16113-16118
Clifton R., Millar A.H., and Whelan J. Alternative oxidases in Arabidopsis: a comparative analysis of differential expression in the gene family provides new insights into function of non-phosphorylating bypasses. Biochim. Biophys. Acta 1757 (2006) 730-741
Ma S., and Bohnert H.J. Integration of Arabidopsis thaliana stress-related transcript profiles, promoter structures, and cell-specific expression. Genome Biol. 8 (2007) R49
Folsch H., Guiard B., Neupert W., and Stuart R.A. Internal targeting signal of the BCS1 protein: a novel mechanism of import into mitochondria. EMBO J. 15 (1996) 479-487
Stan T., Brix J., Schneider-Mergener J., Pfanner N., Neupert W., and Rapaport D. Mitochondrial protein import: recognition of internal import signals of BCS1 by the TOM complex. Mol. Cell. Biol. 23 (2003) 2239-2250
Erzberger J.P., and Berger J.M. Evolutionary relationships and structural mechanisms of AAA+ proteins. Annu. Rev. Biophys. Biomol. Struct. 35 (2006) 93-114
White S.R., and Lauring B. AAA+ ATPases: achieving diversity of function with conserved machinery. Traffic 8 (2007) 1657-1667
Snider J., and Houry W.A. AAA+ proteins: diversity in function, similarity in structure. Biochem. Soc. Trans. 36 (2008) 72-77
Koppen M., and Langer T. Protein degradation within mitochondria: versatile activities of AAA proteases and other peptidases. Crit. Rev. Biochem. Mol. Biol. 42 (2007) 221-242
Lupas A.N., and Martin J. AAA proteins. Curr. Opin. Struct. Biol. 12 (2002) 746-753
Hanson P.I., and Whiteheart S.W. AAA+ proteins: have engine, will work. Nat. Rev. Mol. Cell. Biol. 6 (2005) 519-529
Cruciat C.M., Brunner S., Baumann F., Neupert W., and Stuart R.A. The cytochrome bc1 and cytochrome c oxidase complexes associate to form a single supracomplex in yeast mitochondria. J. Biol. Chem. 275 (2000) 18093-18098
Chi A., Huttenhower C., Geer L.Y., Coon J.J., Syka J.E., Bai D.L., et al. Analysis of phosphorylation sites on proteins from Saccharomyces cerevisiae by electron transfer dissociation (ETD) mass spectrometry. Proc. Natl Acad. Sci. USA 104 (2007) 2193-2198
Ogura T., Whiteheart S.W., and Wilkinson A.J. Conserved arginine residues implicated in ATP hydrolysis, nucleotide-sensing, and inter-subunit interactions in AAA and AAA+ ATPases. J. Struct. Biol. 146 (2004) 106-112
Smith G.R., Contreras-Moreira B., Zhang X., and Bates P.A. A link between sequence conservation and domain motion within the AAA+ family. J. Struct. Biol. 146 (2004) 189-204
Saint-Georges Y., Bonnefoy N., di Rago J.P., Chiron S., and Dujardin G. A pathogenic cytochrome b mutation reveals new interactions between subunits of the mitochondrial bc1 complex. J. Biol. Chem. 277 (2002) 49397-49402
Wach A., Brachat A., Pohlmann R., and Philippsen P. New heterologous modules for classical or PCR-based gene disruptions in Saccharomyces cerevisiae. Yeast 10 (1994) 1793-1808
Brachmann C.B., Davies A., Cost G.J., Caputo E., Li J., Hieter P., and Boeke J.D. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14 (1998) 115-132
Dujardin G., Pajot P., Groudinsky O., and Slonimski P.P. Long range control circuits within mitochondria and between nucleus and mitochondria. I. Methodology and phenomenology of suppressors. Mol. Gen. Genet. 179 (1980) 469-482
Sikorski R.S., and Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122 (1989) 19-27
Longtine M.S., McKenzie III A., Demarini D.J., Shah N.G., Wach A., Brachat A., et al. Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14 (1998) 953-961
Gauss R., Trautwein M., Sommer T., and Spang A. New modules for the repeated internal and N-terminal epitope tagging of genes in Scharomyces cerevisiae. Yeast 22 (2005) 1-12
Kermorgant M., Bonnefoy N., and Dujardin G. Oxa1p, which is required for cytochrome c oxidase and ATP synthase complex formation, is embedded in the mitochondrial inner membrane. Curr. Genet. 31 (1997) 302-307
Schagger H., and Pfeiffer K. Supercomplexes in the respiratory chains of yeast and mammalian mitochondria. EMBO J. 19 (2000) 1777-1783
Lemaire C., and Dujardin G. Preparation of respiratory chain complexes from Saccharomyces cerevisiae wild-type and mutant mitochondria: activity measurement and subunit composition analysis. Methods Mol. Biol. 432 (2008) 65-81
Foury F., and Cazzalini O. Deletion of the yeast homologue of the human gene associated with Friedreich's ataxia elicits iron accumulation in mitochondria. FEBS Lett. 411 (1997) 373-377
Gille C., and Frommel C. STRAP: editor for STRuctural Alignments of Proteins. Bioinformatics 17 (2001) 377-378
Eswar N., Webb B., Marti-Renom M.A., Madhusudhan M.S., Eramian D., Shen M.Y., et al. Comparative protein structure modeling using MODELLER. Curr. Protoc. Protein Sci (2007) Chapter 2, Unit 2 9
Laskowski R.A., Moss D.S., and Thornton J.M. Main-chain bond lengths and bond angles in protein structures. J. Mol. Biol. 231 (1993) 1049-1067
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