title: DNA light-strand preferential recognition of human mitochondria transcription termination factor mTERF.
abstract: Transcription termination of the human mitochondrial genome requires specific binding to termination factor mTERF. In this study, mTERF was produced in E. coli and purified by two-step chromatography. mTERF-binding DNA sequences were isolated from a pool of randomized sequences by the repeated selection of bound sequences by gel-mobility shift assay and polymerase chain reaction. Sequencing and comparison of the 23 isolated clones revealed a 16-bp consensus sequence of 5\'-GTGTGGCA GANCCNGG-3\' in the light-strand (underlined residues were absolutely conserved), which nicely matched the genomic 13-bp terminator sequence 5\'-TGGCAGAGCCC GG-3\'. Moreover, mTERF binding assays of heteroduplex and single-stranded DNAs showed mTERF recognized the light strand in preference to the heavy strand. The preferential binding of mTERF with the light-strand may explain its distinct orientation-dependent termination activity.
authors.affiliation: Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea.
authors.forename: Sang-Chul
authors.initials: SC
authors.lastname: Nam
authors.forename: Changwon
authors.initials: C
authors.lastname: Kang
mesh_terms: D001483:Base Sequence; D050976:Basic-Leucine Zipper Transcription Factors / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D001671:Biochemistry / Q000379:methods; D002845:Chromatography / Q000379:methods; D004247:DNA / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D009154:Mutation; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D014158:Transcription, Genetic
chemical_list: D050976:Basic-Leucine Zipper Transcription Factors; C105178:MTERF1 protein, human; D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; D004247:DNA