Methods to Unravel the Roles of ATPases in Fe-S Cluster Biosynthesis

Iron–sulfur (Fe–S) clusters are present in more than 200 different types of enzymes or proteins and constitute one of the most ancient, ubiquitous and structurally diverse classes of biological prosthetic groups. Hence the process of Fe–S cluster biosynthesis is essential to almost all forms of life and is remarkably conserved in prokaryotic and eukaryotic organisms. Three distinct types of Fe–S cluster assembly machinery have been established in bacteria, termed the NIF, ISC and SUF systems, and, in each case, the overall mechanism involves cysteine desulfurase-mediated assembly of transient clusters on scaffold proteins and subsequent transfer of pre-formed clusters to apo proteins. A molecular level understanding of the complex processes of Fe–S cluster assembly and transfer is now beginning to emerge from the combination of in vivo and in vitro approaches. The present review highlights recent developments in understanding the mechanism of Fe–S cluster assembly and transfer involving the ubiquitous U-type scaffold proteins and the potential roles of accessory proteins such as Nfu proteins and monothiol glutaredoxins in the assembly, storage or transfer of Fe–S clusters.

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Iron–sulfur cluster biosynthesis: characterization of IscU–IscS complex formation and a structural model for sulfide delivery to the [2Fe–2S] assembly site

JBIC Journal of Biological Inorganic Chemistry ◽

10.1007/s00775-009-0495-7 ◽

2009 ◽

Vol 14 (6) ◽

pp. 829-839 ◽

Cited By ~ 10

Author(s):

Manunya Nuth ◽

J. A. Cowan

Keyword(s):

Complex Formation ◽

Structural Model ◽

Iron Sulfur Cluster ◽

Sulfur Cluster ◽

Iron Sulfur ◽

Cluster Biosynthesis ◽

Assembly Site

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Fe-S Cluster Biosynthesis Controls Uptake of Aminoglycosides in a ROS-Less Death Pathway

Science ◽

10.1126/science.1238328 ◽

2013 ◽

Vol 340 (6140) ◽

pp. 1583-1587 ◽

Cited By ~ 129

Author(s):

Benjamin Ezraty ◽

Alexandra Vergnes ◽

Manuel Banzhaf ◽

Yohann Duverger ◽

Allison Huguenot ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Reactive Oxygen ◽

Proton Motive Force ◽

Iron Limitation ◽

Ros Production ◽

Oxygen Species ◽

Fenton Chemistry ◽

Respiratory Complexes ◽

Iron Sulfur ◽

Cluster Biosynthesis

All bactericidal antibiotics were recently proposed to kill by inducing reactive oxygen species (ROS) production, causing destabilization of iron-sulfur (Fe-S) clusters and generating Fenton chemistry. We find that the ROS response is dispensable upon treatment with bactericidal antibiotics. Furthermore, we demonstrate that Fe-S clusters are required for killing only by aminoglycosides. In contrast to cells, using the major Fe-S cluster biosynthesis machinery, ISC, cells using the alternative machinery, SUF, cannot efficiently mature respiratory complexes I and II, resulting in impendence of the proton motive force (PMF), which is required for bactericidal aminoglycoside uptake. Similarly, during iron limitation, cells become intrinsically resistant to aminoglycosides by switching from ISC to SUF and down-regulating both respiratory complexes. We conclude that Fe-S proteins promote aminoglycoside killing by enabling their uptake.

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