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Characterization of RING Finger/SPRY Domain Containing 1 (Rspry1) and NEFA‐interacting Nuclear Protein 30 (Nip30) Transcriptional Regulation
Author(s) -
Duffin Paige,
Waddell David
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.880.13
Subject(s) - ubiquitin ligase , biology , muscle atrophy , reporter gene , myogenesis , microbiology and biotechnology , yy1 , c2c12 , gene , protein degradation , zinc finger , ring finger , gene expression , regulation of gene expression , transcriptional regulation , ubiquitin , myocyte , atrophy , promoter , genetics , transcription factor
Muscle atrophy results from a range of conditions, including immobilization, spinal cord damage, inflammation and aging. An investigation designed to identify changes in gene expression in skeletal muscle following denervation revealed a set of genes that show differential expression patterns in response to neurogenic atrophy. Nip30 and Rspry1, two genes not previously characterized in muscle, were found to be transcriptionally induced during muscle wasting in mice and protein expression was confirmed in C 2 C 12 cells by western blot. In order to characterize the transcriptional regulation of Nip30 and Rspry1, a fragment of the promoter region of each gene was cloned, fused to a reporter gene, and transfected into muscle cells in combination with expression plasmids for Muscle RING Finger 1 (MuRF1) and myogenic regulatory factors (MRFs). The MuRF1 protein is an E3 ubiquitin ligase that is induced under most atrophic conditions and is believed to promote protein degradation; however, data presented in this study suggests that MuRF1 may also regulate the transcriptional activity of genes that are differentially expressed following neurogenic atrophy. Myogenic regulatory factors (MRFs) are a class of E‐box binding proteins that regulate muscle‐specific gene expression. The results of this study demonstrate that MuRF1 and MRFs cooperatively repress Rspry1 and Nip30 reporter activity. Furthermore, mutation of a conserved E‐box element in the shared regulatory region of Nip30 and Rspry1 eliminated the MuRF1 and MRF cooperative repression of these genes. Identifying novel atrophy‐induced genes and characterizing their role in the atrophy cascade may help further our understanding of the molecular mechanisms of muscle wasting.