Expression of the Three Unlinked Isocoding Actin Genes ofPhysarum polycephalum

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Abstract
The actin gene family in Physarum polycephalum contains four unlinked loci: ardA, ardB, ardC, and ardD. The ardA locus is complex and probably contains two genes which we designated ardA2-7 and ardA2-17. cDNA clones corresponding to the ardB and ardC loci were isolated. Nucleic acid sequencing showed that these two cDNAs coded for the only abundant form of Physarum actin, which is 96% homologous to human γ-cytoplasmic actin. The ardA2-17 gene also codes for this same actin protein (Nader et al., Gene 48, 133–144, 1986). The coding regions of ardB and ardC differ by 15 nucleotides. A comparison of the ardB and ardC sequences with ardA2-17 showed 73 and 77 nucleotide substitutions, respectively, in the coding regions. The noncoding regions of these three sequences were not homologous to each other or to the noncoding regions of actin genes from other organisms. Southern genomic hybridizations indicated that the ardA2-7 and ardD genes have weak sequence similarities to the three isocoding actin genes and thus form a different subclass of the family. Northern hybridizations showed that the ardB and ardC transcripts varied in abundance but were present in all the developmental stages. No ardA2-17 transcripts were seen. The relative abundance of the ardB and ardC transcripts was measured in amoebae and plasmodia by S1 nuclease protection and dot hybridization assays. A ratio of approximately 3:1 for ardC versus ardB was found for both stages. P. polycephalum is the first organism shown to contain three unlinked isocoding actin genes, of which at least two are expressed.