Difference between revisions of "PMID:23836870"

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'''Finkenwirth, F, Kirsch, F and Eitinger, T'''  (2013) Solitary BioY proteins mediate biotin transport into recombinant Escherichia coli. ''J. Bacteriol.'' '''195''':4105-11
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!align=left align='left' bgcolor='#CCCCFF' |Abstract
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Energy-coupling factor (ECF) transporters form a large group of vitamin uptake systems in prokaryotes. They are composed of highly diverse, substrate-specific, transmembrane proteins (S units), a ubiquitous transmembrane protein (T unit), and homo- or hetero-oligomeric ABC ATPases. Biotin transporters represent a special case of ECF-type systems. The majority of the biotin-specific S units (BioY) is known or predicted to interact with T units and ABC ATPases. About one-third of BioY proteins, however, are encoded in organisms lacking any recognizable T unit. This finding raises the question of whether these BioYs function as transporters in a solitary state, a feature ascribed to certain BioYs in the past. To address this question in living cells, an Escherichia coli K-12 derivative deficient in biotin synthesis and devoid of its endogenous high-affinity biotin transporter was constructed as a reference strain. This organism is particularly suited for this purpose because components of ECF transporters do not naturally occur in E. coli K-12. The double mutant was viable in media containing either high levels of biotin or a precursor of the downstream biosynthetic path. Importantly, it was nonviable on trace levels of biotin. Eight solitary bioY genes of proteobacterial origin were individually expressed in the reference strain. Each of the BioYs conferred biotin uptake activity on the recombinants, which was inferred from uptake assays with [(3)H]biotin and growth of the cells on trace levels of biotin. The results underscore that solitary BioY transports biotin across the cytoplasmic membrane.
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[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=23836870 PubMed] [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3754743 PMC3754743]
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Online version:[http://dx.doi.org/10.1128/JB.00350-13 10.1128/JB.00350-13]
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!align=left align='left' bgcolor='#CCCCFF' |Keywords
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ATP-Binding Cassette Transporters/genetics; ATP-Binding Cassette Transporters/metabolism; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/metabolism; Biological Transport, Active; Biotin/metabolism; Escherichia coli K12/genetics; Escherichia coli K12/growth & development; Escherichia coli K12/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Membrane Transport Proteins/genetics; Membrane Transport Proteins/metabolism; Mutation; Recombination, Genetic
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==Main Points of the Paper ==
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{{LitSignificance}}
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== Materials and Methods Used ==
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==Phenotype Annotations==
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==Notes==
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==References==
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<references/>
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[[Category:Publication]]

Latest revision as of 18:33, 8 November 2013

Citation

Finkenwirth, F, Kirsch, F and Eitinger, T (2013) Solitary BioY proteins mediate biotin transport into recombinant Escherichia coli. J. Bacteriol. 195:4105-11

Abstract

Energy-coupling factor (ECF) transporters form a large group of vitamin uptake systems in prokaryotes. They are composed of highly diverse, substrate-specific, transmembrane proteins (S units), a ubiquitous transmembrane protein (T unit), and homo- or hetero-oligomeric ABC ATPases. Biotin transporters represent a special case of ECF-type systems. The majority of the biotin-specific S units (BioY) is known or predicted to interact with T units and ABC ATPases. About one-third of BioY proteins, however, are encoded in organisms lacking any recognizable T unit. This finding raises the question of whether these BioYs function as transporters in a solitary state, a feature ascribed to certain BioYs in the past. To address this question in living cells, an Escherichia coli K-12 derivative deficient in biotin synthesis and devoid of its endogenous high-affinity biotin transporter was constructed as a reference strain. This organism is particularly suited for this purpose because components of ECF transporters do not naturally occur in E. coli K-12. The double mutant was viable in media containing either high levels of biotin or a precursor of the downstream biosynthetic path. Importantly, it was nonviable on trace levels of biotin. Eight solitary bioY genes of proteobacterial origin were individually expressed in the reference strain. Each of the BioYs conferred biotin uptake activity on the recombinants, which was inferred from uptake assays with [(3)H]biotin and growth of the cells on trace levels of biotin. The results underscore that solitary BioY transports biotin across the cytoplasmic membrane.

Links

PubMed PMC3754743 Online version:10.1128/JB.00350-13

Keywords

ATP-Binding Cassette Transporters/genetics; ATP-Binding Cassette Transporters/metabolism; Adenosine Triphosphatases/genetics; Adenosine Triphosphatases/metabolism; Biological Transport, Active; Biotin/metabolism; Escherichia coli K12/genetics; Escherichia coli K12/growth & development; Escherichia coli K12/metabolism; Escherichia coli Proteins/genetics; Escherichia coli Proteins/metabolism; Membrane Transport Proteins/genetics; Membrane Transport Proteins/metabolism; Mutation; Recombination, Genetic

Main Points of the Paper

Please summarize the main points of the paper.

Materials and Methods Used

Please list the materials and methods used in this paper (strains, plasmids, antibodies, etc).

Phenotype Annotations

See Help:AnnotationTable for details on how to edit this table.

Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status

Notes

References

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