Crystal structures reveal how distinct sites on the cysteine desulfurase IscS bind two different sulfur-acceptor proteins, IscU and TusA, to transfer sulfur atoms for iron-sulfur cluster biosynthesis and tRNA thiolation

A. D Smith

Allan Matte

Ariane Proteau

B Tirupati

B. W Matthews

C Ayala-Castro

C Pastore

C. D Lima

C. M Wright

C. T Lauhon

C. W Gehrke

D Bordo

D Kessler

D. C Johnson

D. G Waterman

E Katoh

E. C Raulfs

E. G Mueller

F Bou-Abdallah

F Forlani

F Pierrel

G Jager

G Layer

G Musco

G. N Murshudov

Gregory A. Petsko

H Ding

H Li

H Mihara

H. D Urbina

H. K Lundgren

Ismaïl Moukadiri

J. H Kim

J. N Agar

J. R Cupp-Vickery

Jean-François Trempe

K Nilsson

K. A Datsenko

K. G Hoff

L Zheng

L. C Storoni

Linhua Zhang

M Fontecave

M Landau

M Martinez-Vicente

M Nair

M. Eugenia Armengod

Magda Villarroya

Miroslaw Cygler

N. T Perna

P Emsley

P. D Adams

P. M Palenchar

R Kambampati

R Leipuviene

R. A Laskowski

Rong Shi

S Adinolfi

S Dhe-Paganon

S Kato

S. J Cho

T Fujii

T. A Ramelot

U Tokumoto

W Zhang

Y Hu

Y Ikeuchi

Y Shimomura

Z Otwinowski

English

PubMed

Crossref

Structural Basis for Fe–S Cluster Assembly and tRNA Thiolation Mediated by IscS Protein–Protein Interactions

The cysteine desulfurase IscS is a highly conserved master enzyme initiating sulfur transfer via persulfide to a range of acceptor proteins involved in Fe-S cluster assembly, tRNA modifications, and sulfur-containing cofactor biosynthesis. Several IscS-interacting partners including IscU, a scaffold for Fe-S cluster assembly; TusA, the first member of a sulfur relay leading to sulfur incorporation into the wobble uridine of several tRNAs; ThiI, involved in tRNA modification and thiamine biosynthesis; and rhodanese RhdA are sulfur acceptors. Other proteins, such as CyaY/frataxin and IscX, also bind to IscS, but their functional roles are not directly related to sulfur transfer. We have determined the crystal structures of IscS-IscU and IscS-TusA complexes providing the first insight into their different modes of binding and the mechanism of sulfur transfer. Exhaustive mutational analysis of the IscS surface allowed us to map the binding sites of various partner proteins and to determine the functional and biochemical role of selected IscS and TusA residues. IscS interacts with its partners through an extensive surface area centered on the active site Cys328. The structures indicate that the acceptor proteins approach Cys328 from different directions and suggest that the conformational plasticity of a long loop containing this cysteine is essential for the ability of IscS to transfer sulfur to multiple acceptor proteins. The sulfur acceptors can only bind to IscS one at a time, while frataxin and IscX can form a ternary complex with IscU and IscS. Our data support the role of frataxin as an iron donor for IscU to form the Fe-S clusters

M Eugenia Armengod

Directory of Open Access Journals

PLoS Biology

Structural basis for Fe-S cluster assembly and tRNA thiolation mediated by IscS protein-protein interactions.

Shi Rong

Proteau Ariane

Villarroya Magda

Moukadiri Ismaïl

Zhang Linhua

Trempe Jean-François

Matte Allan

Armengod M. Eugenia

Cygler Miroslaw

Public Library of Science (PLOS)

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Role of conserved cysteines in mediating sulfur transfer from IscS to IscU.

sites of disulfide bridge formation in a covalently bound IscS/IscU complex: implications for the mechanism of ironsulfur cluster assembly.

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The cysteine desulfurase IscS is required for synthesis of all five thiolated nucleosides present in tRNA from Salmonella enterica serovar typhimurium.

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Structural Basis for Fe–S Cluster Assembly and tRNA Thiolation Mediated by IscS Protein–Protein Interactions

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