1471-2199-10-96.pdf 2,43MB
1000 Titel
  • Dual effect of a single nucleotide polymorphism in the first intron of the porcine Secreted phosphoprotein 1 gene: allele-specific binding of C/EBP beta and activation of aberrant splicing
1000 Autor/in
  1. Muráni, Eduard |
  2. Ponsuksili, Siriluck |
  3. Seyfert, Hans-Martin |
  4. Shi, Xuanming |
  5. Wimmers, Klaus |
1000 Erscheinungsjahr 2009
1000 Art der Datei
1000 Publikationstyp
  1. Artikel |
1000 Online veröffentlicht
  • 2009-10-21
1000 Erschienen in
1000 Quellenangabe
  • 10: 96
1000 FRL-Sammlung
1000 Copyrightjahr
  • 2009
1000 Lizenz
1000 Verlagsversion
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1000 Ergänzendes Material
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1000 Publikationsstatus
1000 Begutachtungsstatus
1000 Sprache der Publikation
1000 Abstract/Summary
  • BACKGROUND: Secreted phosphoprotein 1 (SPP1 or Osteopontin, OPN) is a multifunctional matricellular glycoprotein involved in development and regeneration of skeletal muscle. Previously, we have demonstrated that porcine SPP1 shows breed-related differential mRNA expression during myogenesis. With the aim to identify putative contributing cis-regulatory DNA variation we resequenced the 5' upstream region of the gene in the respective breeds Pietrain and Duroc. We found two single nucleotide polymorphisms (SNP; [GenBank:M84121]: g.1804C>T and g.3836A>G). We focused our investigation on the SNP g.3836A>G, because in silico analysis and knowledge about the regulation of SPP1 suggested an effect of this SNP on a CCAAT/enhancer binding protein beta (C/EBPβ) responsive transcriptional enhancer. RESULTS: Using electrophoretic mobility shift assay we demonstrated that, similar to human SPP1, the 3' terminal end of the first intron of porcine SPP1 harbors a C/EBPβ binding site and showed that this binding site is negatively affected by the mutant G allele. Genotyping of 48 fetuses per breed revealed that the G allele segregated exclusively in Duroc fetuses with a frequency of 57 percent. Using real-time quantitative PCR we showed that, consistent with its negative effect on a transcriptional enhancer element, the G allele tends to decrease mRNA abundance of SPP1 in the fetal musculus longissimus dorsi (~1.3 fold; P ≥ 0.1). Moreover, we showed that the SNP g.3836A>G leads to ubiquitous aberrant splicing of the first intron by generating a de novo and activating a cryptic splice acceptor site. Aberrantly spliced transcripts comprise about half of the SPP1 messages expressed by the G allele. Both aberrant splice variants differ from the native transcript by insertions in the leader sequences which do not change the reading frame of SPP1. CONCLUSION: At the 3' terminal end of the first intron of the porcine SPP1 we identified a unique, dually functional SNP g.3836A>G. This SNP affects the function of the SPP1 gene at the DNA level by affecting a C/EBPβ binding site and at the RNA level by activating aberrant splicing of the first intron, and thus represents an interesting DNA-marker to study phenotypic effects of SPP1 DNA-variation.
1000 Fachgruppe
  1. Agrarwissenschaften |
1000 Fächerklassifikation (DDC)
1000 Liste der Beteiligten
1000 Förderer
  1. German Research Foundation (Deutsche Forschungsgemeinschaft, DFG; Forschergruppe 'DRIP', FOR 753 |
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    1000 Förderer German Research Foundation (Deutsche Forschungsgemeinschaft, DFG; Forschergruppe 'DRIP', FOR 753 |
    1000 Förderprogramm -
    1000 Fördernummer -
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1000 Erstellt am 2018-01-15T14:27:36.450+0100
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