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Open Access Research article

A genome-wide survey for prion-regulated miRNAs associated with cholesterol homeostasis

Judith Montag1, Markus Brameier2, Ann-Christin Schmädicke1, Sabine Gilch34, Hermann M Schätzl34 and Dirk Motzkus1*

Author Affiliations

1 German Primate Center, Unit of Infection Models, Kellnerweg, 4, 37077, Göttingen, Germany

2 German Primate Center, Primate Genetics Laboratory, Kellnerweg 4, 37077, Göttingen, Germany

3 Institute of Virology, Technical University of Munich (TUM), Trogerstrasse 30, D-81675, Munich, Germany

4 Wyoming Excellence Chair in Prion Biology, Department of Veterinary Sciences and of Molecular Biology, University of Wyoming, 1000 E. University Ave, Laramie, USA

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BMC Genomics 2012, 13:486  doi:10.1186/1471-2164-13-486

Published: 17 September 2012

Abstract

Background

Prion diseases are neurodegenerative diseases that are characterized by the conversion of the cellular prion protein (PrPc) into a pathogenic isoform (PrPSc). It is known that neurodegeneration is often accompanied by the disturbance of cholesterol homeostasis. We have recently identified a set of genes that were upregulated after prion infection of N2a neuronal cells (Bach et al., 2009).

Results

We have now used ultra-deep sequencing technology to profile all microRNAs (miRNA) that could be associated with this effect in these N2a cells. Using stringent filters and normalization strategies we identified a small set of miRNAs that were up- or downregulated upon prion infection. Using bioinformatic tools we predicted whether the downregulated miRNAs could target mRNAs that have been previously identified to enhance cholesterol synthesis in these cells. Application of this joint profiling approach revealed that nine miRNAs potentially target cholesterol-related genes. Four of those miRNAs are localized in a miRNA-dense cluster on the mouse X-chromosome. Among these, twofold downregulation of mmu-miR-351 and mmu-miR-542-5p was confirmed by qRT-PCR. The same miRNAs were predicted as putative regulators of the sterol regulatory element-binding factor 2 (Srebf2), the low-density lipoprotein receptor (Ldlr) or the IPP isomerase.

Conclusions

The results demonstrate that joined profiling by ultra-deep sequencing is highly valuable to identify candidate miRNAs involved in prion-induced dysregulation of cholesterol homeostasis.

Keywords:
Prion disease; Cholesterol; MicroRNA; Ultra-deep sequencing; Joint-profiling