Add time:08/13/2019         Source:sciencedirect.com

SummaryThe Tetrahymena 14-nm filament protein (designated as 49K protein) is a cytoskeletal protein involved in oral morphogenesis and in pronuclear behavior during fertilization. From the previous gene analysis, the 49K protein was shown to share a high sequence identity with porcine heart citrate synthase, and in fact, the 49K protein possessed citrate synthase activity and was detected in mitochondria. These results led to the hypothesis that the 49K protein has dual functions. Since the 49K protein and citrate synthase are clearly different in function and localization, it is necessary to ascertain strictly whether the gene and its transcript are the same or not. We first performed Southern blot analysis in the genomic DNA using three probes, probe 1 corresponding to full length cDNA, probe 2 corresponding to the sequence of 49K protein except for the signal sequence, and probe 3 corresponding to the mitochondrial signal sequence. The Southern hybridizations revealed a reasonable correspondence between the digestion patterns using several restriction enzymes and the patterns deduced from the restriction maps of the genomic 49K protein gene. Neither band suggesting another type of gene was detected in the Southern hybridization under the low stringency conditions. Secondly, Northern hybridizations using the three probes revealed that the transcript of the 49K protein gene was also the same single type. These results indicate that the 14-nm filament protein in the cytoskeleton and citrate synthase in mitochondria are derived from the same gene and the same transcript.

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