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A more recent version of this article appeared on January 1, 2003
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Submitted on June 12, 2002
Revised on September 21, 2002
Accepted on October 3, 2002
1 Department of Biology, Clark University, 950 Main St., Worcester, Massachusetts 01610
* Corresponding author. E-mail address: dlaroche{at}black.clarku.edu.
Here we describe the identification and characterization of the cytokinesis deficient mutant cell line 17HG5 which was generated in a REMI mutagenesis screen designed to isolate genes required for cytokinesis in Dictyostelium discoideum. Phenotypic characterization of the 17HG5 cell line revealed no apparent defects in the global functionality of the actomyosin cytoskeleton except for the observed cytokinesis defect when grown in suspension culture. Plasmid rescue was used to identify the disrupted gene locus (pats1) that caused the cytokinesis defect. Disruption of the pats1 locus was recreated through homologous recombination in several independent cell lines, each recapitulating the cytokinesis-defective phenotype and thereby confirming that this gene locus is important for proper cytokinesis. Sequence data obtained by analysis of the genomic region flanking the inserted REMI plasmid revealed an 8892bp genomic open reading frame encoding a 2964 amino acid protein. The putative pats1 protein contains three regulatory domains (RI-phosphatase, RII- GTP-binding, R-III protein kinase), thirteen leucine-rich repeats, and eight WD-40 repeats. These regulatory domains coupled with the protein-protein interacting domains suggest pats1 is involved in signal transduction during cytokinesis in Dictyostelium.
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