Identification of Genes Differentially Expressed in Myogenin Knock-Down Bovine Muscle Satellite Cells during Differentiation through RNA Sequencing Analysis

Title
Identification of Genes Differentially Expressed in Myogenin Knock-Down Bovine Muscle Satellite Cells during Differentiation through RNA Sequencing Analysis
Author(s)
최인호이은주아델말릭포커렐스미리티사라프라즈아마드아마드미르비랄조경현김지회공준찬[공준찬]이동목[이동목]정기용[정기용]김상훈[김상훈]
Keywords
SKELETAL MYOBLAST DIFFERENTIATION; ENHANCER FACTOR-2 MEF2; STEM-CELLS; TRANSCRIPTION FACTORS; SIGNALING PATHWAY; CYCLE WITHDRAWAL; DNA-REPLICATION; UP-REGULATION; MCM PROTEINS; MYOD
Issue Date
201403
Publisher
PUBLIC LIBRARY SCIENCE
Citation
PLOS ONE, v.9, no.3
Abstract
Background: The expression of myogenic regulatory factors (MRFs) consisting of MyoD, Myf5, myogenin (MyoG) and MRF4 characterizes various phases of skeletal muscle development including myoblast proliferation, cell-cycle exit, cell fusion and the maturation of myotubes to form myofibers. Although it is well known that the function of MyoG cannot be compensated for other MRFs, the molecular mechanism by which MyoG controls muscle cell differentiation is still unclear. Therefore, in this study, RNA-Seq technology was applied to profile changes in gene expression in response to MyoG knockdown (MyoG(kd)) in primary bovine muscle satellite cells (MSCs). Results: About 61-64% of the reads of over 42 million total reads were mapped to more than 13,000 genes in the reference bovine genome. RNA-Seq analysis identified 8,469 unique genes that were differentially expressed in MyoG(kd). Among these genes, 230 were up-regulated and 224 were down-regulated by at least four-fold. DAVID Functional Annotation Cluster (FAC) and pathway analysis of all up- and down-regulated genes identified overrepresentation for cell cycle and division, DNA replication, mitosis, organelle lumen, nucleoplasm and cytosol, phosphate metabolic process, phosphoprotein phosphatase activity, cytoskeleton and cell morphogenesis, signifying the functional implication of these processes and pathways during skeletal muscle development. The RNA-Seq data was validated by real time RT-PCR analysis for eight out of ten genes as well as five marker genes investigated. Conclusions: This study is the first RNA-Seq based gene expression analysis of MyoG(kd) undertaken in primary bovine MSCs. Computational analysis of the differentially expressed genes has identified the significance of genes such as SAP30-like (SAP30L), Protein lyl-1 (LYL1), various matrix metalloproteinases, and several glycogenes in myogenesis. The results of the present study widen our knowledge of the molecular basis of skeletal muscle development and reveal the vital regulatory role of MyoG in retaining muscle cell differentiation.
URI
http://hdl.handle.net/YU.REPOSITORY/32927http://dx.doi.org/10.1371/journal.pone.0092447
ISSN
1932-6203
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생명공학부 > 생명공학부 > Articles
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