Identification, stability and expression of Sirt1 antisense long non-coding RNA

Gene. 2014 Apr 10;539(1):117-24. doi: 10.1016/j.gene.2014.01.037. Epub 2014 Jan 28.

Abstract

Natural antisense transcripts (NATs) exist ubiquitously as pivotal molecules to regulate coding gene expression. Sirtuin 1 (Sirt1) is a NAD-dependent deacetylase which is involved in myogenesis. However, whether Sirt1 transcribes NAT during C2C12 differentiation is still unknown. In this study, we identified a Sirt1 NAT which was designated as Sirt1 antisense long non-coding RNA (AS lncRNA) by sequencing and bioinformatic analysis. The level of Sirt1 AS lncRNA was greater in spleen but less in muscle tissue. The expression of both Sirt1 mRNA and Sirt1 AS lncRNA decreased during C2C12 myogenic differentiation, whereas the levels of miR-34a, which targets Sirt1, increased gradually. We further found that the half-life of Sirt1 AS lncRNA was 10h, but that of Sirt1 mRNA was 6h in C2C12 cells treated with 2 μg/ml Actinomycin D. Therefore, compared with Sirt1 mRNA, Sirt1 AS lncRNA was more stable. Overexpression of Sirt1 AS lncRNA increased the levels of Sirt1 protein, whereas overexpression of Sirt1 AS lncRNA mutant did not affect the level of Sirt1 protein in C2C12 cells. Moreover, downregulation of Sirt1 mRNA caused by miR-34a was counteracted by Sirt1 AS lncRNA in C2C12 cells. Taken together, we identified a novel NAT of Sirt1 which implicated in myogenesis through regulating Sirt1 expression.

Keywords: C2C12; Differentiation; NAT; Sirt1; lncRNA; miR-34a.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Differentiation / genetics
  • Cell Line
  • Dactinomycin / pharmacology
  • Down-Regulation
  • Gene Expression Regulation, Developmental
  • Half-Life
  • Mice
  • MicroRNAs / biosynthesis
  • MicroRNAs / genetics*
  • Molecular Sequence Data
  • Muscle Development / drug effects
  • Muscle Development / genetics*
  • Muscles / cytology
  • RNA Stability / drug effects
  • RNA Stability / genetics
  • RNA, Antisense / biosynthesis
  • RNA, Antisense / genetics*
  • RNA, Long Noncoding / biosynthesis
  • RNA, Long Noncoding / genetics*
  • RNA, Messenger / biosynthesis
  • Sirtuin 1 / genetics*
  • Spleen / cytology

Substances

  • MIRN34a microRNA, mouse
  • MicroRNAs
  • RNA, Antisense
  • RNA, Long Noncoding
  • RNA, Messenger
  • long non-coding RNA SIRT1 AS, mouse
  • Dactinomycin
  • Sirt1 protein, mouse
  • Sirtuin 1