1. RNF19 gene located on human chromosome 8q22.2 has showed 4.4 kb transcript and expressed ubiquitously in various tissues. RNF19 gene coding dorfin protein that carry out E3 ubiquitin ligase, and highly conserved in pig, dog and cattle. RNF19 gene containing MaLR (mammalian LTR-retrotransposon) element in the first intron. Here we found its alternatively spliced transcript variants which derived from MaLR insertion. The MaLR-derived promoter transcripts are detected as two different types in all tissues examined, while breast tissue only showed three variant types. Reporter gene assay of the promoter activity of MaLR element on RNF19 gene indicated good activity in human colon carcinoma cells (HCT- 116). These findings suggest that the MaLR element acquired the role of transcriptional regulation of RNF19 gene in various human tissues during primate evolution.. - cloned from the anterior horn tissues 분류연구 genome transcriptome proteome genome 수준에서의 연구는 이루어지지 않았음 cDNA 서열결정 northern blot 으로 발현패턴 확인 ALS 에서의 전사체 발현양 연구 E3 activity 의 확인 dorfin 이 존재하는 위치확인 신경퇴행성질병과의 관련성 연구 RBRfamily 의 계통연구 parkin 과의 homology 연구 활성메카니즘에 관련된 연구 (VCP 관련 ) CaR 의 degradation 에 미치는 영향 연구 NM_01543 5.3 Parkinson’s disease Amyotrophic lateral sclerosis Other region 16% Gene-related Sequence 36% LINE 20% SINE 13% Coding sequence 3% Pseudogene 1% HERV element 8% DNA element 3% RT-PCR Realtime RT-PCR Cloning & Sequencing Promoter assay Sin HS, Huh JW, Kim DS, Kang DW, Min DS, Kim TH, Ha HS, Kim HH, Lee SY, Kim HS. 2006. Transcriptional control of the HERV-H LTR element of the GSDML gene in human tissues and cancer cells. Arch. Virol. [Epub ahead of print] Hishikawa N, Niwa J, Doyu M, Ito T, Ishigaki S, Hashizume Y, Sobue G. 2003. Dorfin localizes to the ubiquitylated inclusions in Parkinson's disease, dementia with Lewy bodies, multiple system atrophy, and amyotrophic lateral sclerosis. Am J Pathol. 163(2):609-619. Marker Adrenal gland Adult brain Fetal brain Fetal liver Heart Kidney Liver Lung Bone marrow Cerebellum Skeletal muscle Marker Placenta Spinal cord Testis Thymus Thyroid Trachea Uterus Prostate Dorfin Gapdh 494 bp 195 bp 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 MaLR-derived promoter transcript (556 bp) MaLR-derived promoter transcript (430 bp) Relative Expression Bone marrow Cerebellum Kidney Lung Testis Reverse Forward Reverse Forward Control Relative Luciferase Activity (Fold of pGL-2 control) 0 2 4 6 8 10 12 14 16 18 20 22 Cos7 HCT116 CCCCACAAAA GATATGTTCA TGTCCTAATC CCCAGAATCT GCAAATGTTA TTTGGAAAAA GGGGTTTTGC AGATGTAATT AAGTTAAGAA TCTTGAGATA AGATCATCCT GGATTATCCA GGTAGCCTCA AAATCAAGTG ACAAGTGTCT TTGTAAGGGA CAAGTAGACC CATTACAGAG AAGACGACGC GCAGAAAAGG AGGAAGCAGT GTGCTCATGG AGGCGGAGAT TGGAGTGATG TAACCGCAAG CCGAGGAATG CTTATAGTCA CCAGAAGCTG GAAGAGTCAA AGGACACATT CTCCCCTCAA GCCCCAGTGG GAGCACGGCC CAGCTGGATT TTGGACTTCT GGCCTCCAGA ACTGTAAGAG AAATGTCCAT TGTCTTAAGC CAACCAGTTT GTGGTAGTTT GTTACAGCAG CCCCAGGAAA CTACTA. GATA-1 GATA-2 Nkx2-5 WhnELF-1 +1 TRANSCRIPTION START SITE In silico analysis of transcription binding sites of MaLR element 100 77 42 100 Human Dog Pig Cattle Mouse Rat Zebrafish Chicken 35cycle 94 ℃ : 40sec 56 ℃ : 30sec 72 ℃ : 40sec 40cycle (Sybr green) 94 ℃ : 10sec 56 ℃ : 15sec 72 ℃ : 15sec Qiaquick Gel-extraction Promega T-easy Vector EcoR-I(DH5α) Modified PGL2-vector HCT116 Cos7 Lipofectamine Dual luciferase Assay