Identification, stability and expression of Sirt1 antisense long non-coding RNA.
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.
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China. Electronic address: wang_yu@nwsuaf.edu.cn.
Yu
Y
Wang
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China. Electronic address: pwj1226@nwsuaf.edu.cn.
Wei-Jun
WJ
Pang
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Ning
N
Wei
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Yan
Y
Xiong
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Wen-Jing
WJ
Wu
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Cun-Zhen
CZ
Zhao
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Qing-Wu
QW
Shen
College of Animal Science and Technology, Northwest A&F University, Laboratory of Animal Fat Deposition & Muscle Development, Yangling Shaanxi 712100, China.
Gong-She
GS
Yang
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D003609:Dactinomycin / Q000494:pharmacology; D015536:Down-Regulation; D018507:Gene Expression Regulation, Developmental; D006207:Half-Life; D051379:Mice; D035683:MicroRNAs / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D024510:Muscle Development / Q000187:drug effects / Q000235:genetics*; D009132:Muscles / Q000166:cytology; D020871:RNA Stability / Q000187:drug effects / Q000235:genetics; D016372:RNA, Antisense / Q000096:biosynthesis / Q000235:genetics*; D062085:RNA, Long Noncoding / Q000096:biosynthesis / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D056564:Sirtuin 1 / Q000235:genetics*; D013154:Spleen / Q000166:cytology
C551848:MIRN34a microRNA, mouse; D035683:MicroRNAs; D016372:RNA, Antisense; D062085:RNA, Long Noncoding; D012333:RNA, Messenger; C000619513:long non-coding RNA SIRT1 AS, mouse; D003609:Dactinomycin; C470545:Sirt1 protein, mouse; D056564:Sirtuin 1
C2C12; Differentiation; NAT; Sirt1; lncRNA; miR-34a