2.50
Hdl Handle:
http://hdl.handle.net/10033/19238
Title:
NRF IRES activity is mediated by RNA binding protein JKTBP1 and a 14-nt RNA element.
Authors:
Reboll, Marc René; Oumard, André; Gazdag, Aniko Carla; Renger, Isabelle; Ritter, Birgit; Schwarzer, Michael; Hauser, Hansjoerg; Wood, Monika; Yamada, Michiyuki; Resch, Klaus; Nourbakhsh, Mahtab
Abstract:
The mRNA of human NF-kappaB repressing factor (NRF) contains a long 5'-untranslated region (UTR) that directs ribosomes to the downstream start codon by a cap-independent mechanism. Comparison of the nucleotide (nt) sequences of human and mouse NRF mRNAs reveals a high degree of identity throughout a fragment of 150 nt proximal to the start codon. Here, we show that this region constitutes a minimal internal ribosome entry segment (IRES) module. Enzymatic RNA structure analysis reveals a secondary structure model of the NRF IRES module. Point mutation analysis of the module determines a short, 14-nt RNA element (nt 640-653) as a mediator of IRES function. Purification of IRES binding cellular proteins and subsequent ESI/MS/MS sequence analysis led to identification of the RNA-binding protein, JKTBP1. EMSA experiments show that JKTBP1 binds upstream to the 14-nt RNA element in the NRF IRES module (nt 579-639). Over-expression of JKTBP1 significantly enhances activity of the NRF IRES module in dicistronic constructs. Moreover, siRNA experiments demonstrate that down-regulation of endogenous JKTBP1 decreases NRF IRES activity and the level of endogenous NRF protein. The data of this study show that JKTBP1 and the 14-nt element act independently to mediate NRF IRES activity.
Affiliation:
Institute of Pharmacology, Hannover Medical School, Hannover, Germany.
Citation:
NRF IRES activity is mediated by RNA binding protein JKTBP1 and a 14-nt RNA element. 2007, 13 (8):1328-40 RNA
Journal:
RNA (New York, N.Y.)
Issue Date:
Aug-2007
URI:
http://hdl.handle.net/10033/19238
DOI:
10.1261/rna.545407
PubMed ID:
17592041
Additional Links:
http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17592041
Type:
Article
Language:
en
ISSN:
1355-8382
Appears in Collections:
Publications of Dept. Gene Regulation and Differentiation (RDIF)

Full metadata record

DC FieldValue Language
dc.contributor.authorReboll, Marc René-
dc.contributor.authorOumard, André-
dc.contributor.authorGazdag, Aniko Carla-
dc.contributor.authorRenger, Isabelle-
dc.contributor.authorRitter, Birgit-
dc.contributor.authorSchwarzer, Michael-
dc.contributor.authorHauser, Hansjoerg-
dc.contributor.authorWood, Monika-
dc.contributor.authorYamada, Michiyuki-
dc.contributor.authorResch, Klaus-
dc.contributor.authorNourbakhsh, Mahtab-
dc.date.accessioned2008-02-27T14:16:06Z-
dc.date.available2008-02-27T14:16:06Z-
dc.date.issued2007-08-
dc.identifier.citationNRF IRES activity is mediated by RNA binding protein JKTBP1 and a 14-nt RNA element. 2007, 13 (8):1328-40 RNAen
dc.identifier.issn1355-8382-
dc.identifier.pmid17592041-
dc.identifier.doi10.1261/rna.545407-
dc.identifier.urihttp://hdl.handle.net/10033/19238-
dc.description.abstractThe mRNA of human NF-kappaB repressing factor (NRF) contains a long 5'-untranslated region (UTR) that directs ribosomes to the downstream start codon by a cap-independent mechanism. Comparison of the nucleotide (nt) sequences of human and mouse NRF mRNAs reveals a high degree of identity throughout a fragment of 150 nt proximal to the start codon. Here, we show that this region constitutes a minimal internal ribosome entry segment (IRES) module. Enzymatic RNA structure analysis reveals a secondary structure model of the NRF IRES module. Point mutation analysis of the module determines a short, 14-nt RNA element (nt 640-653) as a mediator of IRES function. Purification of IRES binding cellular proteins and subsequent ESI/MS/MS sequence analysis led to identification of the RNA-binding protein, JKTBP1. EMSA experiments show that JKTBP1 binds upstream to the 14-nt RNA element in the NRF IRES module (nt 579-639). Over-expression of JKTBP1 significantly enhances activity of the NRF IRES module in dicistronic constructs. Moreover, siRNA experiments demonstrate that down-regulation of endogenous JKTBP1 decreases NRF IRES activity and the level of endogenous NRF protein. The data of this study show that JKTBP1 and the 14-nt element act independently to mediate NRF IRES activity.en
dc.language.isoenen
dc.relation.urlhttp://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=17592041en
dc.subject.mesh5' Untranslated Regionsen
dc.subject.meshAnimalsen
dc.subject.meshBase Sequenceen
dc.subject.meshDNA-Binding Proteinsen
dc.subject.meshGenes, Reporteren
dc.subject.meshHumansen
dc.subject.meshMiceen
dc.subject.meshMolecular Sequence Dataen
dc.subject.meshNucleic Acid Conformationen
dc.subject.meshProtein Isoformsen
dc.subject.meshRepressor Proteinsen
dc.subject.meshRibonucleoproteinsen
dc.subject.meshRibosomesen
dc.titleNRF IRES activity is mediated by RNA binding protein JKTBP1 and a 14-nt RNA element.en
dc.typeArticleen
dc.contributor.departmentInstitute of Pharmacology, Hannover Medical School, Hannover, Germany.en
dc.identifier.journalRNA (New York, N.Y.)en

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