Antisense RNA inactivation of gene expression of a cell-cell adhesion protein (gp64) in the cellular slime mold Polysphondylium pallidum.
The gp64 protein of Polysphondylium pallidum has been shown to mediate EDTA-stable cell-cell adhesion. To explore the functional role of gp64, we made an antisense RNA expression construct designed to prevent the gene expression of gp64; the construct was introduced into P. pallidum cells and the transformants were characterised. The antisense RNA-expressing clone L3mc2 which had just been harvested at the growth phase tended to re-form in aggregates smaller in size than did the parental cells in either the presence or absence of 10 mM EDTA. In contrast, 6.5-hour starved L3mc2 cells remained considerably dissociated from each other after 5 minutes gyrating, although aggregation gradually increased by 50% during a further 55 minutes gyrating in the presence of 10 mM EDTA. Correspondingly, L3mc2 lacked specifically the cell-cell adhesion protein, gp64. We therefore conclude that the gp64 protein is involved in forming the EDTA-resistant cell-cell contact. In spite of the absence of gp64, L3mc2 exhibited normal developmental processes, a fact which demonstrates that another cell-cell adhesion system exists in the development of Polysphondylium. This is the first report in which an antisense RNA technique was successfully applied to Polysphondylium.
Funamoto
S
S
Division of Biological Sciences, Graduate School of Science, Hokkaido University, Sapporo, Japan.
Ochiai
H
H
D015815:Cell Adhesion Molecules / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005822:Genetic Vectors; D008562:Membrane Glycoproteins / Q000235:genetics*; D009235:Myxomycetes / Q000235:genetics*; D016372:RNA, Antisense / Q000235:genetics*; D014170:Transformation, Genetic; D014764:Viral Proteins
D015815:Cell Adhesion Molecules; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D016372:RNA, Antisense; D014764:Viral Proteins