WhiD is required for the late stages of sporulation in the Gram-positive
bacterium Streptomyces coelicolor. WhiD is a member of the WhiB-like
family of putative transcription factors that are present throughout
the actinomycetes but absent from other organisms. This family of
proteins has four near-invariant cysteines, suggesting that these
residues might act as ligands for a metal cofactor. Overexpressed
WhiD, purified from Escherichia coli, contained substoichiometric
amounts of iron and had an absorption spectrum characteristic of
a 2Fe-2S cluster. After Fe-S cluster reconstitution under anaerobic
conditions, WhiD contained approximately 4 iron atoms/monomer and
similar amounts of sulfide ion and gave an absorption spectrum characteristic
of a 4Fe-4S cluster. Reconstituted WhiD gave no electron paramagnetic
resonance signal as prepared but, after reduction with dithionite,
gave an electron paramagnetic resonance signal (g approximately
2.06, 1.94) consistent with a one-electron reduction of a 4Fe-4S(2+)
cluster to a 4Fe-4S(1+) state with electron spin of S = (1/2).
The anaerobically reconstituted 4Fe-4S cluster was oxygen sensitive.
Upon exposure to air, absorption at 410 and 505 nm first increased
and then showed a steady decrease with time until the protein was
colorless in the near UV/visible region. These changes are consistent
with an oxygen-induced change from a 4Fe-4S to a 2Fe-2S cluster,
followed by complete loss of cluster from the protein. Each of the
four conserved cysteine residues, Cys-23, -53, -56, and -62, was
essential for WhiD function in vivo.
%0 Journal Article
%1 citeulike:519798
%A Jakimowicz, P.
%A Cheesman, M. R.
%A Bishai, W. R.
%A Chater, K. F.
%A Thomson, A. J.
%A Buttner, M. J.
%C Department of Molecular Microbiology, John Innes Centre, Norwich
NR4 7UH, United Kingdom. Piotr.Jakimowicz@bbsrc.ac.uk
%D 2005
%J J Biol Chem
%K iron uv analysis reference
%N 9
%P 8309--8315
%R 10.1074/jbc.M412622200
%T Evidence that the Streptomyces developmental protein WhiD, a member
of the WhiB family, binds a 4Fe-4S cluster.
%U http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=15615709
%V 280
%X WhiD is required for the late stages of sporulation in the Gram-positive
bacterium Streptomyces coelicolor. WhiD is a member of the WhiB-like
family of putative transcription factors that are present throughout
the actinomycetes but absent from other organisms. This family of
proteins has four near-invariant cysteines, suggesting that these
residues might act as ligands for a metal cofactor. Overexpressed
WhiD, purified from Escherichia coli, contained substoichiometric
amounts of iron and had an absorption spectrum characteristic of
a 2Fe-2S cluster. After Fe-S cluster reconstitution under anaerobic
conditions, WhiD contained approximately 4 iron atoms/monomer and
similar amounts of sulfide ion and gave an absorption spectrum characteristic
of a 4Fe-4S cluster. Reconstituted WhiD gave no electron paramagnetic
resonance signal as prepared but, after reduction with dithionite,
gave an electron paramagnetic resonance signal (g approximately
2.06, 1.94) consistent with a one-electron reduction of a 4Fe-4S(2+)
cluster to a 4Fe-4S(1+) state with electron spin of S = (1/2).
The anaerobically reconstituted 4Fe-4S cluster was oxygen sensitive.
Upon exposure to air, absorption at 410 and 505 nm first increased
and then showed a steady decrease with time until the protein was
colorless in the near UV/visible region. These changes are consistent
with an oxygen-induced change from a 4Fe-4S to a 2Fe-2S cluster,
followed by complete loss of cluster from the protein. Each of the
four conserved cysteine residues, Cys-23, -53, -56, and -62, was
essential for WhiD function in vivo.
@article{citeulike:519798,
abstract = {WhiD is required for the late stages of sporulation in the Gram-positive
bacterium Streptomyces coelicolor. WhiD is a member of the WhiB-like
family of putative transcription factors that are present throughout
the actinomycetes but absent from other organisms. This family of
proteins has four near-invariant cysteines, suggesting that these
residues might act as ligands for a metal cofactor. Overexpressed
WhiD, purified from Escherichia coli, contained substoichiometric
amounts of iron and had an absorption spectrum characteristic of
a [2Fe-2S] cluster. After Fe-S cluster reconstitution under anaerobic
conditions, WhiD contained approximately 4 iron atoms/monomer and
similar amounts of sulfide ion and gave an absorption spectrum characteristic
of a [4Fe-4S] cluster. Reconstituted WhiD gave no electron paramagnetic
resonance signal as prepared but, after reduction with dithionite,
gave an electron paramagnetic resonance signal (g approximately
2.06, 1.94) consistent with a one-electron reduction of a [4Fe-4S](2+)
cluster to a [4Fe-4S](1+) state with electron spin of S = (1/2).
The anaerobically reconstituted [4Fe-4S] cluster was oxygen sensitive.
Upon exposure to air, absorption at 410 and 505 nm first increased
and then showed a steady decrease with time until the protein was
colorless in the near UV/visible region. These changes are consistent
with an oxygen-induced change from a [4Fe-4S] to a [2Fe-2S] cluster,
followed by complete loss of cluster from the protein. Each of the
four conserved cysteine residues, Cys-23, -53, -56, and -62, was
essential for WhiD function in vivo.},
added-at = {2007-02-02T11:54:15.000+0100},
address = {Department of Molecular Microbiology, John Innes Centre, Norwich
NR4 7UH, United Kingdom. Piotr.Jakimowicz@bbsrc.ac.uk},
author = {Jakimowicz, P. and Cheesman, M. R. and Bishai, W. R. and Chater, K. F. and Thomson, A. J. and Buttner, M. J.},
biburl = {https://www.bibsonomy.org/bibtex/2c7ef246e854387d8e3f86a73d978d774/robert},
citeulike-article-id = {519798},
comment = {UV Referenzspektren f��r FeS: 420nm},
doi = {10.1074/jbc.M412622200},
interhash = {bdd840c7a10c98c7d616e580563c8ae1},
intrahash = {c7ef246e854387d8e3f86a73d978d774},
issn = {0021-9258},
journal = {J Biol Chem},
keywords = {iron uv analysis reference},
month = {March},
number = 9,
pages = {8309--8315},
priority = {2},
timestamp = {2007-02-02T11:54:15.000+0100},
title = {Evidence that the Streptomyces developmental protein WhiD, a member
of the WhiB family, binds a [4Fe-4S] cluster.},
url = {http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve\&db=pubmed\&dopt=Abstract\&list_uids=15615709},
volume = 280,
year = 2005
}