Cytoimmunofluorescent localization of severin in Dictyostelium amoebae.
Severin is a 40-kDa Ca2+-activated protein from Dictyostelium that rapidly fragments and disassembles actin filaments in vitro (S.S. Brown, K. Yamamoto, and J.A. Spudich, J. Cell Biol. 93, 205-210, 1982; and K. Yamamoto, J.D. Pardee, J. Reidler, L. Stryer, and J.A. Spudich. J. Cell Biol. 95, 711-719, 1982). To determine if severin is colocalized with actin filaments in vivo, we have used the agar-overlay technique of S. Yumura, H. Mori, and Y. Fukui (J. Cell Biol. 99, 894-899, 1984) to examine the intracellular locations of severin and F-actin in vegetative Dictyostelium amoebae. In rounded cells taken from suspension culture severin colocalized with F-actin at cortical edges while maintaining an endoplasmic presence. Both severin and F-actin were present throughout nascent pseudopods of motile cells, while severin appeared concentrated at the leading edge of fully developed pseudopods. Amoebae feeding on a bacterial lawn formed large phagocytic vesicles that were surrounded by an extensive cell cortex rich in severin. Streaming cells entering aggregates during the Dictyostelium developmental cycle showed severin staining throughout the cytoplasm with F-actin at the cortex. The preferential localization of severin in cytoplasmic regions of vegetative cells undergoing extensive actin cytoskeletal rearrangement prompts consideration of a role for severin-mediated disruption of actin filament networks during pseudopod extension and phagocytosis.
Brock
A M
AM
Department of Cell Biology and Anatomy, Cornell University Medical College, New York, New York 10021.
Pardee
J D
JD
D000199:Actins / Q000032:analysis; D000906:Antibodies / Q000276:immunology; D000918:Antibody Specificity; D000941:Antigens / Q000276:immunology; D002448:Cell Adhesion; D002465:Cell Movement; D004023:Dictyostelium / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000032:analysis* / Q000276:immunology; D006651:Histocytochemistry; D008840:Microfilament Proteins / Q000032:analysis* / Q000276:immunology; D015800:Protozoan Proteins
D000199:Actins; D000906:Antibodies; D000941:Antigens; D005656:Fungal Proteins; D008840:Microfilament Proteins; D015800:Protozoan Proteins; C036571:severin protein, Dictyostelium