Screening of genes involved in cell migration in Dictyostelium.
A single cell of wild-type Dictyostelium discoideum forms a visible colony on a plastic dish in several days, but due to enhanced cell migration, amiB-null mutant cells scatter over a large area and do not form noticeable colonies. Here, with an aim to identify genes involved in cell migration, we isolated suppresser mutants of amiB-null mutants that restore the ability to form colonies. From REMI (restriction enzyme-mediated integration)-mutagenized pool of double-mutants, we identified 18 responsible genes from them. These genes can be categorized into several biological processes. One cell line, Sab16 (Suppressor of amiB) was chosen for further analysis, which had a disrupted phospholipase D pldB gene. To confirm the role of pldB gene in cell migration, we knocked out the pldB gene and over-expressed gfp-pldB in wild-type cells. GFP-PLDB localized to plasma membrane and on vesicles, and in migrating cells, at the protruding regions of pseudopodia. Migration speed of vegetative pldB-null cells was reduced to 73% of that of the wild-type. These results suggest that PLDB plays an important role in migration in Dictyostelium cells, and that our screening system is useful for the identification of genes involved in cell migration.
Nagasaki
Akira
A
Research Institute for Cell Engineering, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba Central 4, Tsukuba, Ibaraki, Japan. a-nagasaki@aist.go.jp
Uyeda
Taro Q P
TQ
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002465:Cell Movement / Q000235:genetics*; D022162:Cytoplasmic Vesicles / Q000378:metabolism; D004023:Dictyostelium / Q000235:genetics*; D017125:Genes, Protozoan / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics; D050505:Mutant Proteins; D010739:Phospholipase D / Q000502:physiology*; D021381:Protein Transport; D015800:Protozoan Proteins / Q000235:genetics / Q000502:physiology*; D011554:Pseudopodia / Q000235:genetics
D050505:Mutant Proteins; D015800:Protozoan Proteins; D049452:Green Fluorescent Proteins; D010739:Phospholipase D; C106801:phospholipase D1