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Conference Paper: Mutagenic analysis of the conserved residues of a haloacid permease

TitleMutagenic analysis of the conserved residues of a haloacid permease
Authors
Issue Date2009
PublisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.febsjournal.org/
Citation
The 34th FEBS Congress. Prague, Czech Republic, 4-9 July 2009. In The FEBS Journal, 2009, v. 276 n. S1, p. 168 How to Cite?
AbstractHaloacetic acids are common environmental pollutants and have been shown to be mutagenic. Burkholderia caribensis MBA4, which utilizes monobromoacetate as a growth substrate, was isolated from soil and has been shown to produce an inducible dehalogenase. By means of chromosome walking we have identified a haloacid transporter protein gene (deh4p) downstream of the dehalogenase gene. Comparative analysis showed that Deh4p is a major facilitator superfamily protein, which usually possesses twelve transmembrane segments (TMS). Recombinant DNA technique based on a PhoA-LacZ dual reporters system was used to analyze the topology of Deh4p and the results showed that the N- and the C-termini of the protein were located in the cytoplasm. However, only eight TMS were detected. The first two and the last two putative TMS were not found. ClustalW alignment of Deh4p with some closely related transporter classification TC2.A.1.6 proteins has identified the presence of many conserved residues. An assay system that examined the functional property of the haloacid permease was constructed and used for analysis of mutations on some of these conserved residues. An MBA4 mutant defective in the haloacid operon was used as the host and plasmids with a haloacid operon were transformed into the cell. Charged or potentially phosphorylated residues were selected for modification. A total of sixteen deh4p mutants were generated and tested. The results showed that six of these mutants impeded the growth of the bacterium in utilizing chloroacetate as a growth substrate. Most of these critical mutations were located in the putative cytoplasmic loops of the protein.
Persistent Identifierhttp://hdl.handle.net/10722/114852
ISSN
2015 Impact Factor: 4.237
2015 SCImago Journal Rankings: 2.141

 

DC FieldValueLanguage
dc.contributor.authorTsang, JSH-
dc.contributor.authorTse, YM-
dc.date.accessioned2010-09-26T05:18:50Z-
dc.date.available2010-09-26T05:18:50Z-
dc.date.issued2009-
dc.identifier.citationThe 34th FEBS Congress. Prague, Czech Republic, 4-9 July 2009. In The FEBS Journal, 2009, v. 276 n. S1, p. 168-
dc.identifier.issn1742-464X-
dc.identifier.urihttp://hdl.handle.net/10722/114852-
dc.description.abstractHaloacetic acids are common environmental pollutants and have been shown to be mutagenic. Burkholderia caribensis MBA4, which utilizes monobromoacetate as a growth substrate, was isolated from soil and has been shown to produce an inducible dehalogenase. By means of chromosome walking we have identified a haloacid transporter protein gene (deh4p) downstream of the dehalogenase gene. Comparative analysis showed that Deh4p is a major facilitator superfamily protein, which usually possesses twelve transmembrane segments (TMS). Recombinant DNA technique based on a PhoA-LacZ dual reporters system was used to analyze the topology of Deh4p and the results showed that the N- and the C-termini of the protein were located in the cytoplasm. However, only eight TMS were detected. The first two and the last two putative TMS were not found. ClustalW alignment of Deh4p with some closely related transporter classification TC2.A.1.6 proteins has identified the presence of many conserved residues. An assay system that examined the functional property of the haloacid permease was constructed and used for analysis of mutations on some of these conserved residues. An MBA4 mutant defective in the haloacid operon was used as the host and plasmids with a haloacid operon were transformed into the cell. Charged or potentially phosphorylated residues were selected for modification. A total of sixteen deh4p mutants were generated and tested. The results showed that six of these mutants impeded the growth of the bacterium in utilizing chloroacetate as a growth substrate. Most of these critical mutations were located in the putative cytoplasmic loops of the protein.-
dc.languageeng-
dc.publisherWiley-Blackwell Publishing Ltd.. The Journal's web site is located at http://www.febsjournal.org/-
dc.relation.ispartofThe FEBS Journal-
dc.rightsPreprint This is the pre-peer reviewed version of the following article: [FULL CITE], which has been published in final form at [Link to final article]. Authors are not required to remove preprints posted prior to acceptance of the submitted version. Postprint This is the accepted version of the following article: [full citation], which has been published in final form at [Link to final article].-
dc.titleMutagenic analysis of the conserved residues of a haloacid permease-
dc.typeConference_Paper-
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1742-464X&volume=276 supplement 1&spage=168&epage=&date=2009&atitle=Mutagenic+analysis+of+the+conserved+residues+of+a+haloacid+permease.en_HK
dc.identifier.emailTsang, JSH: jshtsang@hkucc.hku.hk-
dc.identifier.authorityTsang, JSH=rp00792-
dc.identifier.doi10.1111/j.1742-4658.2009.07049.x-
dc.identifier.hkuros168051-
dc.identifier.volume276-
dc.identifier.issueS1-
dc.identifier.spage168-
dc.identifier.epage168-
dc.publisher.placeUnited Kingdom-

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