alpha-Mannosidase-1 mutants of Dictyostelium discoideum: early aggregation-essential genes regulate enzyme precursor synthesis, modification, and processing.
The lysosomal enzyme alpha-mannosidase-1 is one of the earliest developmentally controlled gene products in Dictyostelium discoideum. Although this enzyme is synthesized throughout the first 20 h of development, it is not required for complete morphogenesis, since structural gene (manA) mutants lacking activity develop normally. We isolated six strains deficient in alpha-mannosidase-1 activity which, unlike structural gene mutants, fail to aggregate. Fruiting revertants of these strains accumulate wild-type levels of alpha-mannosidase-1 activity, suggesting that both the enzymatic and morphological defects are caused by single mutations in nonstructural genes essential for early development. Direct genetic evidence for mutations outside of the structural gene was obtained by complementation analysis. We used alpha-mannosidase-1-specific monoclonal antibodies to analyze the biochemical defects in these mad (alpha-mannosidase-1-deficient) mutants. All mad mutants show a significantly reduced relative rate of enzyme precursor biosynthesis. The mad-404 mutation results in a complete lack of precursor biosynthesis, as well as a lack of functional alpha-mannosidase-1 mRNA. In some cases, however, the enzymatic defect results from improper post-translational modification which affects precursor processing. We conclude that a small number of aggregation-essential genes are involved in regulating the synthesis, modification, and processing of alpha-mannosidase-1 during development.
D004023:Dictyostelium / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D004591:Electrophoresis, Polyacrylamide Gel; D004792:Enzyme Precursors / Q000096:biosynthesis* / Q000378:metabolism; D005800:Genes, Fungal; D008361:Mannosidases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D012323:RNA Processing, Post-Transcriptional; D012331:RNA, Fungal / Q000378:metabolism; D013997:Time Factors; D043323:alpha-Mannosidase
D004792:Enzyme Precursors; D012331:RNA, Fungal; D008361:Mannosidases; D043323:alpha-Mannosidase