Friday, March 23, 2012: 8 a.m. - 9:30 a.m.
Presentation Type: Oral Session
MicroRNAs (miRNAs) are small noncoding RNAs that bind to specific mRNA 3’ untranslated regions (UTR) and function as posttranscriptional regulators in development, differentiation and disease. We have identified a 3’UTR alteration in the tooth root master gene NFI-C associated with a novel rare autosomal recessive (AR) form of Radicular Dentin Dysplasia (RDD), a disease characterized by missing roots. Objectives: To characterize the NFI-C 3’UTR and determine the functional role of the AR-RDD alteration on mRNA stability, miRNA regulation and protein level. Methods: AR-RDD and unaffected control dental pulp (DP) cells were grown and harvested for qRT-PCR, immunocytochemistry (ICC), Western blotting (WB), mRNA stability assay, and miRNA PCR array. The 3’UTR region of NFI-C from multiple species was identified (Ensembl) and the degree of conservation determined (ClustalW). Target prediction algorithms (TargetScan, PITA, miRanda, & Micro Inspector) were used to identify potential miRNA binding sites. ICC and WB were performed for NFI-C cellular localization and relative protein levels, respectively. Results: AD-RDD-DP cells were morphologically normal with no alterations in NFI-C cellular localization or isoform expression profile. The identified NFI-C 3’UTR alteration is within a highly conserved element and results in decreased mRNA stability (> 8 hrs) and protein levels (2.2 fold lower). Furthermore, miRNA PCR array data showed differentially expressed miRNAs levels in the AD-RDD-DP cell populations as compared to the control DP. Of the 88 development and differentiation associated miRNAs tested, 15 were downregulated (>2 fold) and 8 were upregulated (> 2 fold) in AR-RDD cells. These miRNAs have targets in genes known to be dysregulated in AR-RDD. Conclusions: Our studies shows for the first time NFI-C is potentially posttranscriptionally regulated by a unique conserved 3’UTR element and that an AR-RDD alteration results in decrease mRNA stability and protein levels resulting in altered signaling cascades critical for root formation. Support: UAB-IOHR/NIDCR-F30DE0180803.
This abstract is based on research that was funded entirely or partially by an outside source: NIDCR-F30DE0180803
Keywords: Gene expression, NFI-C, Pulp, Pulpal disease and Root
See more of: Dentinogenesis: Mechanisms and Therapies
See more of: Pulp Biology & Regeneration Research
See more of: Pulp Biology & Regeneration Research
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