A Novel Cytosolic Isoform of Mitochondrial Trans-2-Enoyl-CoA Reductase Enhances Peroxisome Proliferator-Activated Receptor alpha Activity
Endocrinology and Metabolism
;
: 185-194, 2014.
Article
in English
| WPRIM
| ID: wpr-55019
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Oxidoreductases
/
Yeasts
/
Complement System Proteins
/
Immunohistochemistry
/
HeLa Cells
/
Carrier Proteins
/
Mass Screening
/
Polymerase Chain Reaction
/
Rats, Sprague-Dawley
/
Alternative Splicing
Type of study:
Screening study
Limits:
Animals
/
Humans
Language:
English
Journal:
Endocrinology and Metabolism
Year:
2014
Type:
Article
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