Differential expression and 5' end mapping of actin genes in Dictyostelium

Using a modification of the Berk and Sharp S1 nuclease mapping procedure and by analyzing actin cDNA clones, we have examined the expression of several members of the 17-member multigene family encoding actin in Dictyostelium. The mapping procedure, which takes advantage of the fact that the actin genes are homologous in the protein-coding region but are very divergent in the proposed 5' untranslated region has enabled us to quantitate the relative expression of several genes during the Dictyostelium life cycle. We have shown that at least six of the 17 potential actin-coding sequences are expressed. One is not expressed at levels of more than 0.5--1% of total actin mRNA at the developmental times examined and appears to be a pseudogene. By quantitating the amount of actin mRNA in mRNA populations isolated from cells at various times in development, we have shown that four of the actin genes show different patterns of expression. Interestingly, three of the four genes appear to encode the same protein. We have also taken advantage of the S1 mapping procedure to identify the 5' ends of the actin mRNAs from four genes and have compared the sequences outside the 5' ends on these genes with the nucleotide sequences of seven other actin genes. We have identified homologous sequences in most of these genes that may be involved in initiation of transcription.