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dc.contributor.author
Ramel, Christina
dc.contributor.author
Tobler, Micha
dc.contributor.author
Meyer, Martin
dc.contributor.author
Bigler, Laurent
dc.contributor.author
Ebert, Marc-Olivier
dc.contributor.author
Schellenberg, Barbara
dc.contributor.author
Dudler, Robert
dc.date.accessioned
2019-07-02T09:43:41Z
dc.date.available
2017-06-08T23:37:13Z
dc.date.available
2019-07-02T09:43:41Z
dc.date.issued
2009-10-28
dc.identifier.issn
1472-2091
dc.identifier.other
10.1186/1471-2091-10-26
en_US
dc.identifier.uri
http://hdl.handle.net/20.500.11850/19539
dc.identifier.doi
10.3929/ethz-b-000019539
dc.description.abstract
Background Syringolin A, an important virulence factor in the interaction of the phytopathogenic bacterium Pseudomonas syringae pv. syringae B728a with its host plant Phaseolus vulgaris (bean), was recently shown to irreversibly inhibit eukaryotic proteasomes by a novel mechanism. Syringolin A is synthesized by a mixed non-ribosomal peptide synthetase/polyketide synthetase and consists of a tripeptide part including a twelve-membered ring with an N-terminal valine that is joined to a second valine via a very unusual ureido group. Analysis of sequence and architecture of the syringolin A synthetase gene cluster with the five open reading frames sylA-sylE allowed to formulate a biosynthesis model that explained all structural features of the tripeptide part of syringolin A but left the biosynthesis of the unusual ureido group unaccounted for. Results We have cloned a 22 kb genomic fragment containing the sylA-sylE gene cluster but no other complete gene into the broad host range cosmid pLAFR3. Transfer of the recombinant cosmid into Pseudomonas putida and P. syringae pv. syringae SM was sufficient to direct the biosynthesis of bona fide syringolin A in these heterologous organisms whose genomes do not contain homologous genes. NMR analysis of syringolin A isolated from cultures grown in the presence of NaH13CO3 revealed preferential 13C-labeling at the ureido carbonyl position. Conclusion The results show that no additional syringolin A-specific genes were needed for the biosynthesis of the enigmatic ureido group joining two amino acids. They reveal the source of the ureido carbonyl group to be bicarbonate/carbon dioxide, which we hypothesize is incorporated by carbamylation of valine mediated by the sylC gene product(s). A similar mechanism may also play a role in the biosynthesis of other ureido-group-containing NRPS products known largely from cyanobacteria.
en_US
dc.format
application/pdf
en_US
dc.language.iso
en
en_US
dc.publisher
BioMed Central
en_US
dc.rights.uri
http://creativecommons.org/licenses/by/2.0/
dc.subject
High performance liquid chromatography
en_US
dc.subject
Rice leaves
en_US
dc.subject
Ureido
en_US
dc.subject
Tomato DC3000
en_US
dc.subject
NRPS module
en_US
dc.title
Biosynthesis of the proteasome inhibitor syringolin A: The ureido group joining two amino acids originates from bicarbonate
en_US
dc.type
Journal Article
dc.rights.license
Creative Commons Attribution 2.0 Generic
ethz.journal.title
BMC Biochemistry
ethz.journal.volume
10
en_US
ethz.journal.issue
1
en_US
ethz.journal.abbreviated
BMC biochem.
ethz.pages.start
26
en_US
ethz.size
9 p.
en_US
ethz.version.deposit
publishedVersion
en_US
ethz.identifier.wos
ethz.identifier.nebis
004240256
ethz.publication.place
London
en_US
ethz.publication.status
published
en_US
ethz.leitzahl
08746 - Jaun, Bernhard Mathias (Tit.-Prof.)
en_US
ethz.leitzahl
ETH Zürich::00002 - ETH Zürich::00012 - Lehre und Forschung::00007 - Departemente::02020 - Dep. Chemie und Angewandte Biowiss. / Dep. of Chemistry and Applied Biosc.::02514 - Laboratorium für Organische Chemie / Laboratory of Organic Chemistry::08663 - NMR-Service LOC
en_US
ethz.leitzahl.certified
08746 - Jaun, Bernhard Mathias (Tit.-Prof.)
ethz.date.deposited
2017-06-08T23:37:32Z
ethz.source
ECIT
ethz.identifier.importid
imp59364ca6bcad812532
ethz.ecitpid
pub:31891
ethz.eth
yes
en_US
ethz.availability
Open access
en_US
ethz.rosetta.installDate
2017-07-31T15:15:53Z
ethz.rosetta.lastUpdated
2019-07-02T09:43:51Z
ethz.rosetta.exportRequired
true
ethz.rosetta.versionExported
true
ethz.COinS
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