DNA light-strand preferential recognition of human mitochondria transcription termination factor mTERF.
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.
Nam
Sang-Chul
SC
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 305-701, Korea.
Kang
Changwon
C
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
D050976:Basic-Leucine Zipper Transcription Factors; C105178:MTERF1 protein, human; D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; D004247:DNA