C. Gerez, S. Ollagnier de Choudens, M. Pelosse

Maturation of metallic sites such as iron-sulfur clusters (Fe-S), canonical or complex ones (containing additional heteroatoms), remains a very active and competitive field in bioinorganic chemistry. Maturation systems are structurally and functionally complex and more and more diseases ^[Reference] are revealed due to defects in Fe-S assembly process, pointing to the importance to study at a molecular level such a process.

The BioCatalysis team was pioneer in the study at a molecular level of Fe-S biogenesis in prokaryotes. In bacteria, two conserved Fe-S assembly systems exist, the housekeeping ISC system and the stress-responsive SUF one. Since the late 1990s, the BioCatalysis team is working intensively on the SUF system, composed by six proteins SufA-SufB-SufC-SufD-SufS-SufE, in collaboration with spectroscopists and microbiologists. The team is also working on the ISC system, composed by seven proteins (IscS-IscU-IscA-HscA-HscB-Fdx and CyaY). Currently, a particular attention is focusing on the (i) structural and functional aspects of the SufBCD and SufBCDSE complexes with their cofactors (Fe-S and FADH₂); (ii) iron entry to these complexes to form the Fe-S cluster; (iii) biochemical and biophysical characterization of ISC mutants allowing to bypass frataxin (CyaY), as a strategy to threat Friedreich ataxia in eukaryotes; (iv) biochemical and biophysical characterization of A-Type proteins (SufA, NfuA, IscA, …both wt and variants) and some electron transfer systems allowing an efficient biosynthesis of valuable compounds in cellulo (NAD, Biotin, bisabolene, antibiotics…).

A deep investigation of the eukaryotic system involved in Fe-S biogenesis is essential to better arrest diseases related to Fe-S assembly defects. We are working on the mammalian mitochondrial ISC machinery (Figure 1). Among the twenty proteins involved in this process, we are focusing on (i) ISCA1/2 proteins and its partners involved in the late step of the process and on frataxin (FXN), a protein involved in the initial stage of Fe-S biogenesis and whose deficiency causes the severe neurodegenerative disease Friedreich Ataxia (FRDA).


Figure 1: Mitochondrial ISC machinery.
e-: electron, Fe: iron, apo: apoprotein, X:unknown, CIA: cytosolic Fe-S cluster, red and yellow spheres: iron and sulfur atoms respectively.

Collaborations on the Fe-S biogenesis projects 

F. Barras (Pasteur Institut, Paris); B. Py (LCB, Marseille); H. Puccio (INMG, Lyon); JM. Latour/ G. Blondin (PMB, LCBM, Grenoble); G. Veronesi (MCT, LCBM, Grenoble); J. Pérard (BEE, LCBM, Grenoble); G. Bokinsky (Delf, Netherlands); J.C. Fontecilla-Camps (Métalloprotéines, IBS, Grenoble); S. Cianferani (IPHC, Strasbourg).

Financial supports 

National Agency of Research (ANR FRACOL, ANR EraCoBioTech IronPlugNPlay)

Selected publications on the Fe-S biogenesis project
Py B, Gerez C, Huguenot A, Vidaud C, Fontecave M, de Choudens SO and Barras F
The ErpA/NfuA complex builds an oxidation-resistant Fe-S cluster delivery pathway.
Journal of Biological Chemistry, 2018, 293(20): 7689-7702

Pérard J and Ollagnier de Choudens S
Iron–sulfur clusters biogenesis by the SUF machinery: Close to the molecular mechanism understanding.
Journal of Biological Inorganic Chemistry, 2018, 23(4): 581-596
Correction. Journal of Biological Inorganic Chemistry, 2018, 23(4): 597

Beilschmidt LK, Ollagnier de Choudens S, Fournier M, Sanakis I, Hograindleur MA, Clémancey M, Blondin G, Schmucker S, Eisenmann A, Weiss A, Koebel P, Messaddeq N, Puccio H and Martelli A
ISCA1 is essential for mitochondrial Fe₄S₄ biogenesis in vivo.
Nature Communications, 2017, 8: Article number 15124

Blanc B, Clémancey M, Latour JM, Fontecave M and Ollagnier de Choudens S
Molecular investigation of Iron-Sulfur cluster assembly scaffolds under stress.
Biochemistry, 2014, 53(50): 7867-77869

Colin F, Martelli A, Clemancey M, Latour JM, Gambarelli S, Zeppieri L, Birck C, Page A, Puccio H and Ollagnier de Choudens S
Mammalian frataxin controls sulfur production and iron entry during de novo Fe₄S₄ cluster assembly.
Journal of the American Chemical Society, 2013, 135(2): 733-740

Tsaousis AD, Ollagnier de Choudens S, Gentekaki E, Long S, Gaston D, Stechmann A, Vinella D, Py B, Fontecave M, Barras F, Lukes J and Roger AJ
Evolution of Fe/S cluster biogenesis in the anaerobic parasite Blastocystis.
Proceedings of the National Academy of Sciences, 2012, 109(26): 10426-10431

Wollers S, Layer G, Garcia-Serres R, Signor L, Clemancey M, Latour JM, Fontecave M and Ollagnier de Choudens S
Iron-sulfur (Fe-S) cluster assembly: The SufBCD complex is a new type of Fe-S scaffold with a flavin redox cofactor.
Journal of Biological Chemistry, 2010, 285(30): 23331-23341

Gupta V, Sendra M, Naik SG, Chahal HK, Huynh BH, Outten FW, Fontecave M and Ollagnier de Choudens S
Native Escherichia coli SufA, coexpressed with SufBCDSE, purifies as a [2Fe-2S] protein and acts as an Fe-S transporter to Fe-S target enzymes.
Journal of the American Chemical Society, 2009, 131(17): 6149-6153

Loiseau L, Gerez C, Bekker M, Ollagnier-de Choudens S, Py B, Sanakis Y, Teixeira de Mattos J, Fontecave M and Barras F
ErpA, an iron sulfur (Fe S) protein of the A-type essential for respiratory metabolism in Escherichia coli.
Proceedings of the National Academy of Sciences USA, 2007, 104(34): 13626-13631

Ranquet C, Ollagnier de Choudens S, Loiseau L, Barras F and Fontecave M
Cobalt stress in Escherichia coli: The effect on the iron-sulfur proteins.
Journal of Biological Chemistry, 2007, 282(42): 30442-30451

Layer G, Ollagnier-de Choudens S, Sanakis Y and Fontecave M
Iron-sulfur cluster biosynthesis: characterization of Escherichia coli CYaY as an iron donor for the assembly of [2Fe-2S] clusters in the scaffold IscU.
Journal of Biological Chemistry, 2006, 281(24): 16256-63

Loiseau L, Ollagnier de Choudens S, Fontecave M and Barras F
Analysis of the Escherichia coli CsdA-CsdE heterodimeric cysteine desulfurase, a new [Fe-S] biogenesis system.
Journal of Biological Chemistry, 2005, 280: 26760-26769

Loiseau L, Ollagnier de Choudens S, Nachin L, Fontecave M and Barras F
Biogenesis of Fe-S cluster by the bacterial Suf system: SufS and SufE form a new type of cysteine desulfurase.
Journal of Biological Chemistry, 2003, 278: 28352-28359

Ollagnier de Choudens S, Sanakis Y, Loiseau L, Nachin L, Barras F and Fontecave M
SufA from Erwinia chrysanthemi: Characterization of a scaffold protein required for iron-sulfur cluster assembly.
Journal of Biological Chemistry, 2003, 278: 17993-18001

Ollagnier de Choudens S, Mattioli T, Takahashi Y and Fontecave M
Iron-sulfur cluster assembly: Characterization of IscA and evidence for a specific and functional complex with ferredoxin.
Journal of Biological Chemistry, 2001, 276: 22604-22607


Books
Puccio H and Ollagnier de Choudens S
Iron-sulfur cluster assembly in bacteria and eukarya using the ISC biosynthesis machinery Encyclopedia of Inorganic and Bioinorganic Chemistry, 2017 In ‘Metalloprotein Active Site Assembly’, edited by Michael K. Johnson and Robert A. Scott. Chichester, UK: John Wiley & Sons, Ltd, pp. 1-19.

Barras F and Ollagnier de Choudens S
Genetic, biochemical and biophysical methods for studying Fe-S proteins and their assembly Meth. Enzymol. 2017, Vol 595, Edited by Sheila S. David, pp. 1-32.