Biogenesis of [Fe-S] proteins

1
Biogenesis of Fe-S proteins
Frédéric Barras, LCB, CNRS, Marseille
2
Some Fe-S clusters
Kiley and Beinert, 2003
3
iscAnif
nifS
NIF
nifU
iscR
iscS
iscU
fdx
hscB
iscA
hscA
ISC
sufE
sufS
sufA
sufB
sufC
sufD
SUF
csdA
ygdK
CSD
4
Three machineries in E. coli
iscR
iscS
iscU
fdx
hscB
iscA
hscA
ISC
sufE
sufS
sufA
sufB
sufC
sufD
SUF
csdA
ygdK
CSD
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Cysteine
S donor
Fe
S acceptor
Alanine
S
Fe-S
Scaffold
Fe-S
Target
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SUF machinery
Cysteine
S donor
Fe
S acceptor
Alanine
S
Fe-S
Scaffold
ABC ATPase
?
Fe-S
Target
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ISC machinery
Cysteine
S donor
S acceptor
Fe
Chaperone
Fe-S
Alanine
Fe-S
Scaffold
Ferredoxin
?
Fe-S
Target
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CSD machinery
Cysteine
S donor
Fe
S acceptor
Alanine
S
Fe-S
Target
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Substrate specificity
Suf
Isc
Iron assimilation
Thionucleosides
Fe-S
Thiamin Molybdopterin
Csd
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Environmental specificity
Suf
Isc
H2O2
OxyR
IscR 2Fe-2S
Fur
Csd
Nachin et al., 2001,, Schwartz et al., 2001,
Zheng et al., 2001, Hantke, 2002 Lee et al., 2004
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Biogenesis of Fe-S proteins in Cenibacterium
Phenotypic complementation
E. coli iscS and sufS mutants Library
Cenibacterium
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iscS of Cenibacterium
Complementation thiamine and nicotinic acid
requirement
Cloned DNA
Genome analysis
iscR
iscS
iscU
hscB
iscA
fdx
hscA
52
69
82
67
39
59
72
Genetic organisation identical to E. coli
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Characterisation of IscSCeni
Purification (His)6-IscSCeni
IscSCeni
Cysteine desulfurase activity PLP/protein
µmol alanine/min-1 mig-1 IscScoli
0.22 1 IscSCeni 0.33 1 CsdAcoli
0.5 1
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No SUF machinery in Cenibacterium
Complementation no sufS-containing insert
Genome analysis one cysteine desulfurase IscS
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E. coli sufS mutant complementation (Growth in
presence of iron chelator )
Cloned DNA
orf1
cysA
Carboxynorspermidine dehydrogenase
Peptidase
sodC
cysW
cysT
47 id. Cbl 45 id. CysB
Sulfur metabolism regulator
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(No Transcript)
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One machinery in Cenibacterium
iscR
iscS
iscU
hscB
iscA
fdx
hscA
ISC
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One machinery in Cenibacterium
iscR
iscS
iscU
hscB
iscA
fdx
hscA
ISC
Implication of the ISC machinery in arsenic
resistance
Model to study Scaffold proteins
Problems to construct mutants. iscS, iscA, msrA
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Frédéric Barras Béatrice Py Gabriel Rincón
Enríquez
Collaborators
CEA, Grenoble Sandrine Ollagnier de Choudens Marc
Fontecave