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PMID: 7876307 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Genetic deletion of ABP-120 alters the three-dimensional organization of actin filaments in Dictyostelium pseudopods.

The Journal of cell biology ·Vol. 128 ·No. 5 ·1995-03-00 ·Pages 819-35

Cox D, Ridsdale JA, Condeelis J, Hartwig J

Abstract

This study extends the observations on the defects in pseudopod formation of ABP-120+ and ABP-120- cells by a detailed morphological and biochemical analysis of the actin based cytoskeleton. Both ABP-120+ and ABP-120- cells polymerize the same amount of F-actin in response to stimulation with cAMP. However, unlike ABP-120+ cells, ABP-120- cells do not incorporate actin into the Triton X-100-insoluble cytoskeleton at 30-50 s, the time when ABP-120 is incorporated into the cytoskeleton and when pseudopods are extended after cAMP stimulation in wild-type cells. By confocal and electron microscopy, pseudopods extended by ABP-120- cells are not as large or thick as those produced by ABP-120+ cells and in the electron microscope, an altered filament network is found in pseudopods of ABP-120- cells when compared to pseudopods of ABP-120+ cells. The actin filaments found in areas of pseudopods in ABP-120+ cells either before or after stimulation were long, straight, and arranged into space filling orthogonal networks. Protrusions of ABP-120- cells are less three-dimensional, denser, and filled with multiple foci of aggregated filaments consistent with collapse of the filament network due to the absence of ABP-120-mediated cross-linking activity. The different organization of actin filaments may account for the diminished size of protrusions observed in living and fixed ABP-120- cells compared to ABP-120+ cells and is consistent with the role of ABP-120 in regulating pseudopod extension through its cross-linking of actin filaments.

MeSH Terms
Actin Cytoskeleton/ultrastructure Actins/metabolism Amanitins/metabolism Animals Carrier Proteins/genetics Cyclic AMP/pharmacology Cytoskeleton/drug effects,ultrastructure Dictyostelium/drug effects,genetics,ultrastructure Gene Deletion Microfilament Proteins/genetics Microscopy, Confocal Microscopy, Electron Microscopy, Fluorescence Models, Biological Phagocytes/ultrastructure Polyethylene Glycols/pharmacology Pseudopodia/ultrastructure
Chemicals
Actins Amanitins Carrier Proteins Microfilament Proteins abpC protein, Dictyostelium Polyethylene Glycols 7-nitrobenz-2-oxa-1,3-diazole-phallacidin Cyclic AMP
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Cox D
Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, New York 10461.
Ridsdale J A
Condeelis J
Hartwig J
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.128.5.819
Published
1995-03-00
Pages
819-35
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2120398
Subset
IM
Grants
NIGMS NIH HHS · 5T32 GM25813 · United States
NIGMS NIH HHS · GM25813 · United States
NHLBI NIH HHS · HL47874 · United States
External Links
PubMed source DOI: 10.1083/jcb.128.5.819 PMC Full Text View journal details
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