A Novel Cytosolic Isoform of Mitochondrial Trans-2-Enoyl-CoA Reductase Enhances Peroxisome Proliferator-Activated Receptor alpha Activity.
10.3803/EnM.2014.29.2.185
- Author:
Dong Gyu KIM
1
;
Jae Cheal YOO
;
Eunju KIM
;
Young Sun LEE
;
Oleg V YARISHKIN
;
Da Yong LEE
;
Kun Ho LEE
;
Seong Geun HONG
;
Eun Mi HWANG
;
Jae Yong PARK
Author Information
1. Department of Physiology, Institute of Health Science, Medical Research Center for Neural Dysfunction, Gyeongsang National University School of Medicine, Jinju, Korea. jaeyong@gnu.ac.kr
- Publication Type:Original Article
- Keywords:
Trans-2-enoyl-CoA reductase (NADPH);
Cytosolic mitochondrial trans-2-enoyl-CoA reductase;
PPAR alpha;
Alternative splicing;
Mitochondrial targeting sequences
- MeSH:
Alternative Splicing;
Amino Acids;
Animals;
Carrier Proteins;
Complement System Proteins;
Cytoplasm;
Cytosol*;
Fatty Acids;
Fluorescence;
HeLa Cells;
Humans;
Immunohistochemistry;
Luciferases;
Mass Screening;
Mitochondria;
Oxidoreductases*;
Polymerase Chain Reaction;
PPAR alpha*;
Rats;
Rats, Sprague-Dawley;
Reverse Transcription;
Yeasts
- From:Endocrinology and Metabolism
2014;29(2):185-194
- CountryRepublic of Korea
- Language:English
-
Abstract:
BACKGROUND: Mitochondrial trans-2-enoyl-CoA reductase (MECR) is involved in mitochondrial synthesis of fatty acids and is highly expressed in mitochondria. MECR is also known as nuclear receptor binding factor-1, which was originally reported with yeast two-hybrid screening as a binding protein of the nuclear hormone receptor peroxisome proliferator-activated receptor alpha (PPARalpha). However, MECR and PPARalpha are localized at different compartment, mitochondria, and the nucleus, respectively. Therefore, the presence of a cytosolic or nuclear isoform of MECR is necessary for functional interaction between MECR and PPARalpha. METHODS: To identify the expression pattern of MECR and the cytosolic form of MECR (cMECR), we performed reverse transcription polymerase chain reaction (RT-PCR) with various tissue samples from Sprague-Dawley rats. To confirm the interaction between cMECR and PPARalpha, we performed several binding assays such as yeast two-hybrid, coimmunoprecipitation, and bimolecular fluorescence complementation. To observe subcellular localization of these proteins, immunocytochemistry was performed. A luciferase assay was used to measure PPARalpha activity. RESULTS: We provide evidence of an alternatively spliced variant of the rat MECR gene that yields cMECR. The cMECR lacks the N-terminal 76 amino acids of MECR and shows uniform distribution in the cytoplasm and nucleus of HeLa cells. cMECR directly bound PPARalpha in the nucleus and increased PPARalpha-dependent luciferase activity in HeLa cells. CONCLUSION: We found the cytosolic form of MECR (cMECR) was expressed in the cytosolic and/or nuclear region, directly binds with PPARalpha, and enhances PPARalpha activity.
