Identification of Drosophila cytoskeletal proteins by induction of abnormal cell shape in fission yeast

• Published in: Proceedings of the National Academy of Sciences - PNAS 1994-05, Vol.91 (10), p.4589-4593
• Main Authors: Edwards, K.A, Montague, R.A, Shepard, S, Edgar, B.A, Erikson, R.L, Kiehart, D.P
• Format: Article
• Language: English
• Subjects: actin; Actin Depolymerizing Factors; Actins; Amino Acid Sequence; animal proteins; Animals; binding proteins; cell biology; Cells; Cloning, Molecular - methods; cofilin; Complementary DNA; Conserved Sequence; Cytoskeletal Proteins - biosynthesis; Cytoskeletal Proteins - genetics; Cytoskeletal Proteins - physiology; cytoskeleton; Drosophila; Drosophila melanogaster; Drosophila melanogaster - genetics; Drosophila melanogaster - physiology; ezrin; fungal anatomy; Gene Library; genetic transformation; Genetic Vectors; Humans; Microfilament Proteins - biosynthesis; Microfilament Proteins - genetics; Microfilament Proteins - physiology; Microfilaments; Molecular Sequence Data; Nerve Tissue Proteins - biosynthesis; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - physiology; phenotype; Phenotypes; profilin; Profilins; Proteins; Saccharomycetales; schizosaccharomyce pombe; Schizosaccharomyces - genetics; Schizosaccharomyces pombe; Sequence Homology, Amino Acid; Vertebrates; Yeasts
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.91.10.4589

To clone metazoan genes encoding regulators of cell shape, we have developed a functional assay for proteins that affect the morphology of a simple organism, the fission yeast Schizosaccharomyces pombe. A Drosophila melanogaster cDNA library was constructed in an inducible expression vector and transformed into S. pombe. When expression of the Drosophila sequences was induced, aberrant cell shapes were found in 0.2% of the transformed colonies. Four severe phenotypes representing defects in cytokinesis and/or cell shape maintenance were examined further. Each displayed drastic and specific reorganizations of the actin cytoskeleton. Three of the cDNAs responsible for these defects appear to encode cytoskeletal components: the actin binding proteins profilin and cofilin/actin depolymerizing factor and a membrane-cytoskeleton linker of the ezrin/merlin family. These results demonstrate that a yeast phenotypic screen efficiently identifies conserved genes from more complex organisms and sheds light on their potential in vivo functions.