Box C/D small nucleolar RNA (snoRNA) U60 regulates intracellular cholesterol trafficking

Katrina A. Brandis, Sarah Gale, Sarah Jinn, Stephen J. Langmade, Nicole Dudley-Rucker, Hui Jiang, Rohini Sidhu, Aileen Ren, Anna Goldberg, Jean E. Schaffer, Daniel S. Ory

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Mobilization of plasmamembrane (PM) cholesterol to the endoplasmic reticulum is essential for cellular cholesterol homeostasis. The mechanisms regulating this retrograde, intermembrane cholesterol transfer are not well understood. Because mutant cells with defects in PM to endoplasmic reticulum cholesterol trafficking can be isolated on the basis of resistance to amphotericin B, we conducted an amphotericin B loss-of-function screen in Chinese hamster ovary (CHO) cells using insertional mutagenesis to identify genes that regulate this trafficking mechanism. Mutant line A1 displayed reduced cholesteryl ester formationfrom PM-derived cholesterol and increased de novo cholesterol synthesis, indicating a deficiency in retrograde cholesterol transport. Genotypic analysis revealed that the A1 cell line contained one disrupted allele of the U60 small nucleolar RNA (snoRNA) host gene, resulting in haploinsufficiency of the box C/D snoRNA U60. Complementation and mutational studies revealed the U60 snoRNA to be the essential feature from this locus that affects cholesterol trafficking. Lack of alteration in predicted U60-mediated site-directed methylation of 28 S rRNA in the A1 mutant suggests that the U60 snoRNA modulates cholesterol trafficking by a mechanism that is independent of this canonical function. Our study adds to a growing body of evidence for participation of small noncoding RNAs in cholesterol homeostasis and is the first to implicate a snoRNA in this cellular function.

Original languageEnglish
Pages (from-to)35703-35713
Number of pages11
JournalJournal of Biological Chemistry
Volume288
Issue number50
DOIs
StatePublished - Dec 13 2013

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    Brandis, K. A., Gale, S., Jinn, S., Langmade, S. J., Dudley-Rucker, N., Jiang, H., Sidhu, R., Ren, A., Goldberg, A., Schaffer, J. E., & Ory, D. S. (2013). Box C/D small nucleolar RNA (snoRNA) U60 regulates intracellular cholesterol trafficking. Journal of Biological Chemistry, 288(50), 35703-35713. https://doi.org/10.1074/jbc.M113.488577