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PMID: 10908675 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The cysteine desulfurase, IscS, has a major role in in vivo Fe-S cluster formation in Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 97 ·No. 16 ·2000-08-01 ·Pages 9009-14

Schwartz CJ, Djaman O, Imlay JA, Kiley PJ

Abstract

The cysteine desulfurase, IscS, provides sulfur for Fe-S cluster synthesis in vitro, but a role for IscS in in vivo Fe-S cluster formation has yet to be established. To study the in vivo function of IscS in Escherichia coli, a strain lacking IscS was constructed and characterized. Using this iscS deletion strain, we have observed decreased specific activities for proteins containing [4Fe-4S] clusters from soluble (aconitase B, 6-phosphogluconate dehydratase, glutamate synthase, fumarase A, and FNR) and membrane-bound proteins (NADH dehydrogenase I and succinate dehydrogenase). A specific role for IscS in in vivo Fe-S cluster assembly was demonstrated by showing that an Fe-S cluster independent mutant of FNR is unaffected by the lack of IscS. These data support the conclusion that, via its cysteine desulfurase activity, IscS provides the sulfur that subsequently becomes incorporated during in vivo Fe-S cluster synthesis. We also have characterized a growth phenotype associated with the loss of IscS. Under aerobic conditions the deletion of IscS caused an auxotrophy for thiamine and nicotinic acid, whereas under anaerobic conditions, only nicotinic acid was required. The lack of IscS also had a general effect on the growth of E. coli because the iscS deletion strain grew at half the rate of wild type in many types of media even when the auxotrophies were satisfied.

MeSH Terms
Carbon-Sulfur Lyases/genetics,metabolism Escherichia coli/genetics,growth & development,metabolism Genetic Complementation Test Iron-Sulfur Proteins/biosynthesis,chemistry Mutation Niacin/metabolism Phenotype Thiamine/metabolism
Chemicals
Iron-Sulfur Proteins Niacin Carbon-Sulfur Lyases cysteine desulfurase Thiamine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Schwartz C J
Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI 53706, USA.
Djaman O
Imlay J A
Kiley P J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.160261497
Published
2000-08-01
Pages
9009-14
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC16812
Subset
IM
Grants
NIGMS NIH HHS · R37 GM049640 · United States
NIGMS NIH HHS · GM-45844 · United States
NIGMS NIH HHS · R01 GM045844 · United States
NIGMS NIH HHS · GM49640 · United States
NIGMS NIH HHS · R01 GM049640 · United States
External Links
PubMed source DOI: 10.1073/pnas.160261497 PMC Full Text View journal details
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