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Discrimintion and Mapping of the Primary and Processed Transcripts in Maize Mitochondrion Using a Circular RT-PCR-based Strategy
Published on: July 29, 2019
A mitochondrial gene involved in cytochrome c maturation (ccmC) is expressed as a precursor with a long NH2-terminal
Kazuyoshi Kitazaki1, Yuta Nomoto, Akihiro Aoshima
1Laboratory of Genetic Engineering, Research Faculty of Agriculture, Hokkaido University, Kita-ku, Sapporo 060-8589, Japan.
Abstract:
Extensive genome rearrangement is one of the major mechanisms of angiosperm mitochondrial evolution. As a by-product, some angiosperm mitochondrial genes exhibit divergent organization, but not all of these genes have been fully characterized. Sugar beet ccmC, which plays an important role in cytochrome c maturation, harbors a unique extended NH(2) terminal region of 277 amino acid residues (N-extension) instead of a conserved translational initiation codon. The 5' termini of two major RNA species were determined by primer extension analysis, which revealed that the larger transcript covered the entire N-extension. Nucleotide sequencing of the cDNA revealed that a total of 31 C-to-U RNA editing events occurred in the N-extension and the ccmC-homologous region (ccmC-core region), resulting in improvement of amino acid sequence conservation. Antiserum was raised against a synthetic peptide corresponding to the ccmC-core region and was used for protein gel blot analysis of sugar beet and radish mitochondrial proteins. The detected 29.5-kDa signal band is shared by sugar beet and radish. Two additional larger signal bands are exclusively detected from sugar beet. The largest signal band is also detected by anti-N-extension antiserum. Our results indicate that sugar beet ccmC is translated as a long precursor with N-extension.
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