Sher T, Yi H F, McBride O W, Gonzalez F J
Laboratory of Molecular Carcinogenesis, National Cancer Institute, Bethesda, Maryland 20892.
Biochemistry. 1993 Jun 1;32(21):5598-604. doi: 10.1021/bi00072a015.
The human peroxisome proliferator activated receptor (hPPAR) was cloned from a human liver cDNA library. The cDNA exhibited 85% and 91% DNA and deduced amino acid sequence identity with mouse PPAR (mPPAR), respectively. The hPPAR gene was mapped on human chromosome 22 slightly telomeric to a linkage group of six genes and genetic markers that are located in the general region 22q12-q13.1. Cotransfection assays of mouse Hepa 1 cells were used to roughly compare the ability of hPPAR- and mPPAR-expressed cDNAs to trans-activate the acyl CoA oxidase (ACO) PPAR response element located 5' upstream to the minimal thymidine kinase promoter driving the expression of the chloramphenicol acetyl transferase (CAT) reporter gene. Both receptors elicited a response with the prototypical peroxisome proliferators nafenopin, clofibrate, and WY-14,643. Moreover, using cotransfection assays in which the CAT reporter plasmid contained the CYP4 A6 gene response element rather than the ACO element, it was shown that hPPAR is capable of very efficiently trans-activating a second PPAR response element. These results indicate that the PPAR is present in humans in a form that is functional and can trans-activate response elements derived from two different genes, the rat ACO and the rabbit CYP4A6.
人类过氧化物酶体增殖物激活受体(hPPAR)是从人类肝脏cDNA文库中克隆出来的。该cDNA与小鼠PPAR(mPPAR)的DNA和推导的氨基酸序列一致性分别为85%和91%。hPPAR基因定位于人类22号染色体上,略位于端粒方向,与位于22q12 - q13.1一般区域的六个基因和遗传标记的连锁群相邻。使用小鼠Hepa 1细胞的共转染试验来大致比较表达hPPAR和mPPAR的cDNA对位于驱动氯霉素乙酰转移酶(CAT)报告基因表达的最小胸苷激酶启动子5'上游的酰基辅酶A氧化酶(ACO)PPAR反应元件进行反式激活的能力。两种受体都对典型的过氧化物酶体增殖剂萘芬诺平、氯贝丁酯和WY - 14,643产生反应。此外,使用共转染试验,其中CAT报告质粒包含CYP4 A6基因反应元件而非ACO元件,结果表明hPPAR能够非常有效地反式激活第二个PPAR反应元件。这些结果表明,PPAR在人类中以一种功能性形式存在,并且能够反式激活源自两个不同基因(大鼠ACO和兔CYP4A6)的反应元件。