dc.contributor.author
Graf, Dominik
dc.contributor.author
Wesslowski, Janine
dc.contributor.author
Ma, Hongju
dc.contributor.author
Scheerer, Patrick
dc.contributor.author
Krauß, Norbert
dc.contributor.author
Oberpichler, Inga
dc.contributor.author
Zhang, Fan
dc.contributor.author
Lamparter, Tilman
dc.date.accessioned
2018-06-08T03:12:30Z
dc.date.available
2015-11-20T09:44:57.276Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/14688
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-18879
dc.description.abstract
Photolyases can repair pyrimidine dimers on the DNA that are formed during UV
irradiation. PhrB from Agrobacterium fabrum represents a new group of
prokaryotic (6–4) photolyases which contain an iron-sulfur cluster and a DMRL
chromophore. We performed site-directed mutagenesis in order to assess the
role of particular amino acid residues in photorepair and photoreduction,
during which the FAD chromophore converts from the oxidized to the
enzymatically active, reduced form. Our study showed that Trp342 and Trp390
serve as electron transmitters. In the H366A mutant repair activity was lost,
which points to a significant role of His366 in the protonation of the lesion,
as discussed for the homolog in eukaryotic (6–4) photolyases. Mutants on
cysteines that coordinate the Fe-S cluster of PhrB were either insoluble or
not expressed. The same result was found for proteins with a truncated
C-terminus, in which one of the Fe-S binding cysteines was mutated and for
expression in minimal medium with limited Fe concentrations. We therefore
assume that the Fe-S cluster is required for protein stability. We further
mutated conserved tyrosines that are located between the DNA lesion and the
Fe-S cluster. Mutagenesis results showed that Tyr424 was essential for lesion
binding and repair, and Tyr430 was required for efficient repair. The results
point to an important function of highly conserved tyrosines in prokaryotic
(6–4) photolyases.
en
dc.rights.uri
http://creativecommons.org/licenses/by/4.0/
dc.subject.ddc
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit
dc.title
Key Amino Acids in the Bacterial (6-4) Photolyase PhrB from Agrobacterium
fabrum
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation
PLoS ONE. - 10 (2015), 10, Artikel Nr. e0140955
dcterms.bibliographicCitation.doi
10.1371/journal.pone.0140955
dcterms.bibliographicCitation.url
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0140955
refubium.affiliation
Charité - Universitätsmedizin Berlin
de
refubium.mycore.fudocsId
FUDOCS_document_000000023489
refubium.note.author
Der Artikel wurde in einer Open-Access-Zeitschrift publiziert.
refubium.resourceType.isindependentpub
no
refubium.mycore.derivateId
FUDOCS_derivate_000000005678
dcterms.accessRights.openaire
open access