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Article: Functional studies of the small subunit of EcoHK31I DNA methyltransferase

TitleFunctional studies of the small subunit of EcoHK31I DNA methyltransferase
Authors
KeywordsEMSA
Escherichia coli
Methyltransferase
Protein-DNA interaction
Restriction-modification system
Issue Date2006
PublisherWalter de Gruyter GmbH & Co KG. The Journal's web site is located at http://www.degruyter.de/journals/bc
Citation
Biological Chemistry, 2006, v. 387 n. 5, p. 507-513 How to Cite?
AbstractEcoHK31I DNA methyltransferase recognizes the sequence 5′-YGGCCR- 3′ and adds a methyl group to the fifth position of the internal cytosine to protect the DNA from cleavage by its cognate endonuclease. M.EcoHK31I is composed of polypeptides α and β. Polypeptide β only contains the conserved IX motif of the C5-MTase family, and provides a unique example to show that this motif alone may be dislocated to another polypeptide. By electromobility shift assay, we found that the α/β complex recognizes specific oligonucleotide substrates. Polypeptide α formed aggregates with DNA, while polypeptide β alone did not bind DNA. Therefore, polypeptide β assists in the proper binding of polypeptide α to DNA substrate. The complex of polypeptide α and a polypeptide β variant with an N-terminal deletion of 41 amino acids showed a 16-fold reduction in methylation activity. Further deletion resulted in an inactive methyltransferase. The dissociation equilibrium constant (Kd) of the α/β complex was 56.4 nM, while the Kd value for the α/ΔN46- polypeptide β complex was increased approximately 95-fold, caused by a drastic decrease in dissociate rate constant (kd) and an increase in the association rate constant (ka). This indicates that the N-terminal region of polypeptide β takes part in subunit interaction, while the C-terminal region is involved in DNA binding. Copyright © by Walter de Gruyter.
Persistent Identifierhttp://hdl.handle.net/10722/157447
ISSN
2015 Impact Factor: 2.71
2015 SCImago Journal Rankings: 1.607
ISI Accession Number ID
References

 

DC FieldValueLanguage
dc.contributor.authorFung, WTen_HK
dc.contributor.authorSze, KHen_HK
dc.contributor.authorLee, KFen_HK
dc.contributor.authorShaw, PCen_HK
dc.date.accessioned2012-08-08T08:50:03Z-
dc.date.available2012-08-08T08:50:03Z-
dc.date.issued2006en_HK
dc.identifier.citationBiological Chemistry, 2006, v. 387 n. 5, p. 507-513en_HK
dc.identifier.issn1431-6730en_HK
dc.identifier.urihttp://hdl.handle.net/10722/157447-
dc.description.abstractEcoHK31I DNA methyltransferase recognizes the sequence 5′-YGGCCR- 3′ and adds a methyl group to the fifth position of the internal cytosine to protect the DNA from cleavage by its cognate endonuclease. M.EcoHK31I is composed of polypeptides α and β. Polypeptide β only contains the conserved IX motif of the C5-MTase family, and provides a unique example to show that this motif alone may be dislocated to another polypeptide. By electromobility shift assay, we found that the α/β complex recognizes specific oligonucleotide substrates. Polypeptide α formed aggregates with DNA, while polypeptide β alone did not bind DNA. Therefore, polypeptide β assists in the proper binding of polypeptide α to DNA substrate. The complex of polypeptide α and a polypeptide β variant with an N-terminal deletion of 41 amino acids showed a 16-fold reduction in methylation activity. Further deletion resulted in an inactive methyltransferase. The dissociation equilibrium constant (Kd) of the α/β complex was 56.4 nM, while the Kd value for the α/ΔN46- polypeptide β complex was increased approximately 95-fold, caused by a drastic decrease in dissociate rate constant (kd) and an increase in the association rate constant (ka). This indicates that the N-terminal region of polypeptide β takes part in subunit interaction, while the C-terminal region is involved in DNA binding. Copyright © by Walter de Gruyter.en_HK
dc.languageengen_US
dc.publisherWalter de Gruyter GmbH & Co KG. The Journal's web site is located at http://www.degruyter.de/journals/bcen_HK
dc.relation.ispartofBiological Chemistryen_HK
dc.subjectEMSAen_HK
dc.subjectEscherichia colien_HK
dc.subjectMethyltransferaseen_HK
dc.subjectProtein-DNA interactionen_HK
dc.subjectRestriction-modification systemen_HK
dc.subject.meshBase Sequenceen_US
dc.subject.meshCircular Dichroismen_US
dc.subject.meshDna-Cytosine Methylases - Chemistry - Genetics - Metabolismen_US
dc.subject.meshElectrophoretic Mobility Shift Assayen_US
dc.subject.meshEscherichia Coli - Enzymologyen_US
dc.subject.meshEscherichia Coli Proteins - Chemistry - Genetics - Metabolismen_US
dc.subject.meshProtein Structure, Quaternaryen_US
dc.subject.meshProtein Structure, Secondaryen_US
dc.subject.meshProtein Subunits - Chemistry - Genetics - Metabolismen_US
dc.subject.meshRecombinant Proteins - Biosynthesis - Metabolismen_US
dc.subject.meshSequence Deletion - Geneticsen_US
dc.subject.meshSurface Plasmon Resonanceen_US
dc.titleFunctional studies of the small subunit of EcoHK31I DNA methyltransferaseen_HK
dc.typeArticleen_HK
dc.identifier.emailSze, KH:khsze@hku.hken_HK
dc.identifier.emailLee, KF:ckflee@hku.hken_HK
dc.identifier.authoritySze, KH=rp00785en_HK
dc.identifier.authorityLee, KF=rp00458en_HK
dc.description.naturelink_to_subscribed_fulltexten_US
dc.identifier.doi10.1515/BC.2006.066en_HK
dc.identifier.pmid16740121-
dc.identifier.scopuseid_2-s2.0-33746071515en_HK
dc.identifier.hkuros123135-
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-33746071515&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume387en_HK
dc.identifier.issue5en_HK
dc.identifier.spage507en_HK
dc.identifier.epage513en_HK
dc.identifier.isiWOS:000238135100003-
dc.publisher.placeGermanyen_HK
dc.identifier.scopusauthoridFung, WT=14041658700en_HK
dc.identifier.scopusauthoridSze, KH=7006735061en_HK
dc.identifier.scopusauthoridLee, KF=26643097500en_HK
dc.identifier.scopusauthoridShaw, PC=35599523600en_HK

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