Molecular cloning and characterization of the translationally controlled tumor protein gene in Bombyx mori

• DOI: 10.1016/j.cbpc.2004.06.004
• Publication Date: 2004-08-05T19:44:01Z
• Authors: Jae Man Lee; Takahiro Kusakabe; Yutaka Kawaguchi; Yoshitaka Miyagawa; Masateru Takahashi; Hiroaki Mon; Si-Kab Nho; Katsumi Koga
• Keywords: Expressed Sequence Tags; 0301 basic medicine; DNA, Complementary; Base Sequence; Transcription, Genetic; Molecular Sequence Data; Sequence Homology; Tumor Protein, Translationally-Controlled 1; Genomics; Bombyx; 03 medical and health sciences; Gamma Rays; Biomarkers, Tumor; Animals; Insect Proteins; Amino Acid Sequence; RNA, Messenger; Cloning, Molecular; Promoter Regions, Genetic; Phylogeny

Translationally controlled tumor protein (Tctp/p23) is known to be synthesized preferentially in cells during the early growth phase of tumors, but is also expressed in normal cells. To elucidate its molecular basis of the expression and physiological significance, a cDNA encoding for the Bombyx mori Tctp (BmTctp) was deduced by editing the partial cDNA sequences registered in a Bombyx EST database. RT-PCR analyses indicated that the BmTCTP mRNA was transcribed in all larval organs examined and was present constantly during the cell cycle of BmN4 cells. A genomic clone of 4255 nucloetide residues produced by inverse PCR contained the 5'-flanking region, two introns and three exons of the BmTCTP gene. Sequence analysis of the 5'-flanking region indicated that a putative promoter region contains several canonical transcription elements such as GATA box, CCAAT motif, MEF2, E4BP4.01 and AP-1, but lacks a TATA box element. Luciferase reporter assay of the deletion constructs of the 5'-flanking region revealed that the -676 to +66 region enhanced the promoter activity the most markedly. In addition to this, there were at least two enhancer-like elements and several repressor elements.