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Kim, Min Young,Soh, Jae Mog 한국유전학회 1996 Genes & Genomics Vol.18 No.4
A complementary DNA clone was screened with a single-copy genomic DNA fragment derived from yeast artificial chromosome (YAC) clone D142H8 which contained phosphoribosylglycinamide formyltransferase (GART) gene and an accessory factor-1 (AF-1) gene of human interferon receptor-gamma receptor. The length of the clone is 1337 nt and has an the longest open reading frame (ORF) of 349 amino acids. Genbank search revealed that it is the forth member of quinone oxidoreductase super family. Therefore, the gene was named as quinone oxidoreductase family-4 (qorf-4). The putative protein contains the conserved glycine-rich NADPH binding motif (A/GXGXXGXXXA), which suggest that it might have an enzymatic activity, possibly reductase with NADPH as a cofactor. Multiple tissue Northern blot data showed that the mRNA was expressed ubiquitously. The gene of qorf-4 is located on human chromosome 21q22.1 region where the YAC was mapped. The location of the gene is further localized on the YAC D142H8 with respect to 2 NotI sites and GART and AF-1 loci.
LEE, HYE KYUNG,Kwon, Hyuk Bang,Soh, Jae Mog 한국유전학회 1996 Genes & Genomics Vol.18 No.4
The expression of steroidogenic acute regulatory (StAR) protein is up-regulated in steroidogenic cells in acute response to trophic hormones and the protein is responsible for the mobilization of cholesterol from mitochondrial outer membrane to inner membrane where cholesterol side chain cleavage enzyme reside. Rat complementary DNA corresponding the StAR protein was cloned and the complete nucleotide sequence was determined. The cDNA is consisted of 1560 nucleotides and has the longest open reading frame of 320 amino acids. The deduced amino acids sequence show high degree of homology to mouse, bovine, and human StAR protein in amino acid level. Rat tissue Northern blot showed that it is only expressed in testis, ovary, and adrenal gland. Rat Leydig tumor cells, R2C can produce testosterone without trophic hormone stimulation or cAMP analog treatment. In order to correlate the constitutive production of the hormone with the the level of StAR mRNA, we analyzed the expression of the StAR mRNA in rat R2C cells. The Northern blot showed that R2C cells expressed the StAR mRNA without stimulation and treatment of cAMP did not increase the level of StAR mRNA, which indicates that constitutive steroidogenesis are caused by high basal level expression of StAR mRNA and StAR expression in R2C cells is not dependent on cAMP level.
Characterization and Mapping of Orphan Nuclear Hormone Receptor SHP Gene
LEE, HYE KYUNG,Choi, Hueng Sik,Kwon, Hyuk Bang,Soh, Jae Mog 한국유전학회 1996 Genes & Genomics Vol.18 No.4
We have previously reported that the orphan nuclear hormone receptor SHP act as a heterodimeric partner of several nuclear hormone receptors and it inhibits the activity of these receptors. In order to study the gene regulation mechanism, we have cloned the genomic DNA of SHP from mouse and human genomic library. Exon-intron structure of mouse and human SHP genomic DNA are determined. Both genomic structure contain two exons and one intron. 2.3 kb of mouse and 1.3 kb of human SHP gene. Several potential transcription factor binding sites such as SP-1, AP-1, GATA, ROR, and NF_(κ)B exist in mouse SHP promoter region. SHP gene is localized in human chromosome number 1 by somatic cell hybrid panel and fluoresence in situ hybridization (FISH) analysis.
LEE, HYE KYUNG,Choi, Hueng Sik,Kwon, Hyuk Bang,Soh, Jae Mog,Yoo, Myung Sik 한국유전학회 1996 Genes & Genomics Vol.18 No.4
Retinoic acids (RAs) exert pleiotrophic effects on cellular growth, differentiation, and testicular functions. The primary function of Leydig cells is to produce testosterone whose intratesticular level is critical for spermatogenesis. Steroidogenic Acute Regulatory (StAR) protein is involved in transporting cholesterol from outer mitochondrial membrane to matrix side of inner mitochondrial membrane where cholesterol side chain cleavage (SCC) enzyme reside. Thus, the StAR protein is essential for acute response of steroidogenesis in steroidogenic tissues and the action of the protein is thought to be rate-limiting step for the testosterone synthesis. In order to understand how RAs control steroidogenesis in Leydig cells, cultured mouse Leydig tumor cells K-28 were treated with RAS and the mRNA levels of StAR gene were monitored by Northern blot analysis. Our study showed that RAs up-regulate the StAR mRNA in K-28 cells in time- and dose-dependent manner while the levels of the SCC mRNA were unchanged. The amount of progesterone produced from RA treated K-28 cells was increased concomitantly with the level of StAR mRNA, which indicates that StAR protein is the key regulator for steroidogenesis.