The 25 kDa protein recognizing the rat curved region upstream of the origin of the L-strand replication is the rat homologue of the human mitochondrial transcription factor A.
Mass spectrometry matrix-assisted laser desorption ionization (MALDI) analysis and N-terminus sequencing as well as immunoblotting experiments using human and mouse antibodies have allowed us to identify the 25 kDa protein, previously isolated from rat liver using magnetic beads coated with a rat liver mitochondrial (mt) DNA region upstream of the Ori-L, as the homologue of human mt transcription factor A (mtTFA). We can therefore identify this DNA binding protein as the rat mtTFA. Furthermore, since we previously showed that the 25 kDa protein purified from rat liver was able to bind the curved mtDNA region upstream of the Ori-L as well as the curved mtDNA in the D-loop region, the results here reported lead us to state, for the first time, that mtTFA binds both the curved regions of mtDNA upstream of the two replication origins.
Pierro
P
P
Department of Biochemistry and Molecular Biology, University of Bari, Via Orabona 4, 70125, Bari, Italy.
Capaccio
L
L
Gadaleta
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D004272:DNA, Mitochondrial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000737:chemistry; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D051381:Rats; D018741:Replication Origin; D016415:Sequence Alignment / Q000379:methods; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D029867:Xenopus Proteins
D004272:DNA, Mitochondrial; D004268:DNA-Binding Proteins; D015534:Trans-Activators; C097078:XL-MTTFA protein, Xenopus; D029867:Xenopus Proteins