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Article: Basonuclin regulates a subset of ribosomal RNA genes in HaCAT cells

TitleBasonuclin regulates a subset of ribosomal RNA genes in HaCAT cells
Authors
Advisors
Issue Date2007
PublisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
Citation
Plos One, 2007, v. 2 n. 9 How to Cite?
AbstractBasonuclin (Bnc1), a cell-type specific ribosomal RNA (rRNA) gene regulator, is expressed mainly in keratinocytes of stratified epithelium and gametogenic cells of testis and ovary. Previously, basonuclin was shown in vitro to interact with rRNA gene (rDNA) promoter at three highly conserved sites, Basonuclin's high affinity binding site overlaps with the binding site of a dedicated and ubiquitous Pol I transcription regulator, UBF, suggesting that their binding might interfere with each other if they bind to the same promoter. Knocking-down basonuclin in mouse oocytes eliminated approximately one quarter of RNA polymerase I (Pol I) transcription foci, without affecting the BrU incorporation of the remaining ones, suggesting that basonuclin might regulate a subset of rDNA. Here we show, via chromatin immunoprecipitation (ChIP), that basonuclin is associated with rDNA promoters in HaCaT cells, a spontaneously established human keratinocyte line. Immunoprecipitation data suggestthat basonuclin is in a complexthat also contains the subunits of Pol I (RPA194, RPA116), but not UBF. Knocking down basonuclin in HaCaT cells partially impairs the association of RPA194 to rDNA promoter, but not that of UBF. Basonuclin-deficiency also reduces the amount of 47S pre-rRNA, but this effect can be seen only after cell-proliferation related rRNA synthesis has subsided at a higher cell density. DNA sequence of basonuclin-bound rDNA promoters shows single nucleotide polymorphisms (SNPs) that differ from those associated with UBF-bound promoters suggesting that basonuclin and UBF interact with different subsets of promoters. In conclusion, our results demonstrate basonuclin's functional association with rDNA promoters and its interaction with Pol I in vivo. Our data also suggest that basonuclin-Pol I complex transcribes a subset of rDNA. © 2007 Zhang et al.
Persistent Identifierhttp://hdl.handle.net/10722/147570
ISSN
2015 Impact Factor: 3.057
2015 SCImago Journal Rankings: 1.395
ISI Accession Number ID
Funding AgencyGrant Number
NIHAG14456
Funding Information:

This work was supported by NIH grant AG14456 to HT. The funding agency played no role in the design, conduct and report of this study.

References

 

DC FieldValueLanguage
dc.contributor.advisorCreative Commons: Attribution 3.0 Hong Kong License-
dc.contributor.authorZhang, Sen_US
dc.contributor.authorWang, Jen_US
dc.contributor.authorTseng, Hen_US
dc.date.accessioned2012-05-29T06:04:41Z-
dc.date.available2012-05-29T06:04:41Z-
dc.date.issued2007en_US
dc.identifier.citationPlos One, 2007, v. 2 n. 9en_US
dc.identifier.issn1932-6203en_US
dc.identifier.urihttp://hdl.handle.net/10722/147570-
dc.description.abstractBasonuclin (Bnc1), a cell-type specific ribosomal RNA (rRNA) gene regulator, is expressed mainly in keratinocytes of stratified epithelium and gametogenic cells of testis and ovary. Previously, basonuclin was shown in vitro to interact with rRNA gene (rDNA) promoter at three highly conserved sites, Basonuclin's high affinity binding site overlaps with the binding site of a dedicated and ubiquitous Pol I transcription regulator, UBF, suggesting that their binding might interfere with each other if they bind to the same promoter. Knocking-down basonuclin in mouse oocytes eliminated approximately one quarter of RNA polymerase I (Pol I) transcription foci, without affecting the BrU incorporation of the remaining ones, suggesting that basonuclin might regulate a subset of rDNA. Here we show, via chromatin immunoprecipitation (ChIP), that basonuclin is associated with rDNA promoters in HaCaT cells, a spontaneously established human keratinocyte line. Immunoprecipitation data suggestthat basonuclin is in a complexthat also contains the subunits of Pol I (RPA194, RPA116), but not UBF. Knocking down basonuclin in HaCaT cells partially impairs the association of RPA194 to rDNA promoter, but not that of UBF. Basonuclin-deficiency also reduces the amount of 47S pre-rRNA, but this effect can be seen only after cell-proliferation related rRNA synthesis has subsided at a higher cell density. DNA sequence of basonuclin-bound rDNA promoters shows single nucleotide polymorphisms (SNPs) that differ from those associated with UBF-bound promoters suggesting that basonuclin and UBF interact with different subsets of promoters. In conclusion, our results demonstrate basonuclin's functional association with rDNA promoters and its interaction with Pol I in vivo. Our data also suggest that basonuclin-Pol I complex transcribes a subset of rDNA. © 2007 Zhang et al.en_US
dc.languageengen_US
dc.publisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.actionen_US
dc.relation.ispartofPLoS ONEen_US
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshBase Sequenceen_US
dc.subject.meshBlotting, Northernen_US
dc.subject.meshBlotting, Westernen_US
dc.subject.meshCell Lineen_US
dc.subject.meshChromatin Immunoprecipitationen_US
dc.subject.meshDna Methylationen_US
dc.subject.meshDna-Binding Proteins - Genetics - Physiologyen_US
dc.subject.meshGene Expression Regulationen_US
dc.subject.meshGene Knockdown Techniquesen_US
dc.subject.meshHumansen_US
dc.subject.meshImmunohistochemistryen_US
dc.subject.meshMolecular Sequence Dataen_US
dc.subject.meshPolymorphism, Single Nucleotideen_US
dc.subject.meshPromoter Regions, Geneticen_US
dc.subject.meshRna, Ribosomal - Geneticsen_US
dc.subject.meshSequence Homology, Nucleic Aciden_US
dc.subject.meshTranscription Factors - Genetics - Physiologyen_US
dc.titleBasonuclin regulates a subset of ribosomal RNA genes in HaCAT cellsen_US
dc.typeArticleen_US
dc.identifier.emailWang, J:junwen@hkucc.hku.hken_US
dc.identifier.authorityWang, J=rp00280en_US
dc.description.naturepublished_or_final_versionen_US
dc.identifier.doi10.1371/journal.pone.0000902en_US
dc.identifier.pmid17878937-
dc.identifier.scopuseid_2-s2.0-38549146003en_US
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-38549146003&selection=ref&src=s&origin=recordpageen_US
dc.identifier.volume2en_US
dc.identifier.issue9en_US
dc.identifier.isiWOS:000207455700010-
dc.publisher.placeUnited Statesen_US
dc.identifier.scopusauthoridZhang, S=7409376616en_US
dc.identifier.scopusauthoridWang, J=8950599500en_US
dc.identifier.scopusauthoridTseng, H=19637691100en_US

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