ALTERNATIVE TRANSCIPTION OF HUMAN CHM GENE INCLUDING LTR12C ELEMENT IN COLORECTAL CANCER

1. Receptor binding proteins Lipid envelope RNase Viral RNA Capsid : core proteins TSD LTR gag pol env LTR TSD ALTERNATIVE TRANSCIPTION OF HUMAN CHM GENE INCLUDING LTR12C ELEMENT IN COLORECTAL CANCER Yi-Deun Jung1, Yun-Ji Kim1, Jae-Won Huh2, Dae-Soo Kim3, Hong-Seok Ha1, Eun-Sang Choi1, Won-Ho Lee1 and Heui-Soo Kim1 1 Division of Biological Sciences, College of Natural Sciences, Pusan National University, Busan 609-735, Republic of Korea 2 National Primate Research Center, KRIBB, Ochang, Chungbuk 363-883, Republic of Korea 3 Korea Bioinformation Center, KRIBB, Daejeon 305-806, Republic of Korea purewonder@pusan.ac.kr • Abstract CHM (Choroideremia) is indispensable for the posttranslational activation of retina-specific Rab protein. Various mutations such as missense, nonsense, rearrangement of the CHM gene are responsible for choroideremia. We could find several elements including MIR, LINE, Alu and LTR12C that are located within the intron 4 of the CHM gene. Alternatively spliced transcript including LTR12C element was detected in the last exon of gene. In order to detect this transcript, we accompanied RT-PCR analysis in the normal and adjacent cancer tissue samples, indicating that this transcript was expressed higher in colorectal cancer than normal tissue samples with similar expression patterns in other cancer tissues (liver, lung, testes). We also confirmed high expression of colorectal cancer in patient sample. The expression level of alternative transcript of CHM gene could be used to predict colorectal cancer. Further study about effect of LTR12C in this alternative transcript between normal and cancer tissues is indicated. • Introduction • Materials & Methods • Human & Primate genomic DNA Bioinformatics ( in silico analysis CHM gene ) 8% Genomic DNA PCR & Gene Cloning LTR : long terminal repeat LINE : long integrated nuclear element SINE : short integrated nuclear element HERV : Human endogenous retro-virus RT PCR ( Human original & EST form) colorectal tissue samples RT-PCR, Image J analysis PBS gag : coding for matrix and capsid proteins pol : reverse transcriptase and integrase proteins env : product viral envelope and transmembrane structures • Results & Discussion • CHM ( choroideremia ) • Analyzed quantitatively using the Image-J program Chr X q21.2 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 4 5 85105713 85105326 85120551 85120427 • RT-PCR (original transcript & EST form) M 1 2 3 4 5 6 7 8 9 10 • Alternative splicing transcript EST Original 1 2 3 4 5 M 11 12 13 14 15 16 17 18 19 20 EST • Transcript including LTR12C RT-PCR (colorectal cancer tissue) Original N C N C N C N C N C N C 1 : adrenal gland, 2 : bone marrow, 3 : adult brain cerebellum, 4 : adult brain whole, 5 : fetal brain, 6 : fetal liver, 7 : heart, 8 : kidney, 9 : liver, 10 : lung, 11 : placenta, 12 : salivary gland, 13 : SK-muscle, 14 : spinal cord, 15 : testis, 16 : prostate, 17 : thymus, 18 : thyroid, 19 : trachea, 20 : uterus UTR CDS MIR/SINE Alu/SINE L1/LINE ERV1/LTR trigger5 N C N C N C N C • References • Patience C, Takeuchi Y, Weiss RA (1997) Infection of human cells by an endogenous retrovirus of pigs. Nat Med 3: 276-282. • Scheef G, Fischer N, Krach U, Tonjes RR (2001) The number of a U3 repeat box acting as an enhancer in long terminal repeats of polytropic • replication-competent porcine endogenous retroviruses dynamically fluctuates during serial virus passages in human cells. J Virol 75: 6933-6940