TY - JOUR
T1 - Involvement of profilin in the actin-based motility of L. monocytogenes in cells and in cell-free extracts
AU - Theriot, Julie A.
AU - Rosenblatt, Jody
AU - Portnoy, Daniel A.
AU - Goldschmidt-Clermont, Pascal J.
AU - Mitchison, Timothy J.
N1 - Funding Information:
We would like to thank Lisa Belmont for advice on the preparation of Xenopus egg extracts, Linda Wordeman and Rebecca Bernat for the gift of the HeLa cell line, and Christine Kocks and Pascale Cossart for communication of unpublished data. This work was supported by fellowships from the Packard Foundation and the Searle Foundation (T. J. M.), the Howard Hughes Medical Institute (J. A. T.), and the National Science Foundation (J. R.).
PY - 1994/2/11
Y1 - 1994/2/11
N2 - Within hours of Listeria monocytogenes infection, host cell actin filaments form a dense cloud around the intracytoplasmic bacteria and then rearrange to form a polarized comet tail that is associated with moving bacteria. We have devised a cell-free extract system capable of faithfully reconstituting L. monocytogenes motility, and we have used this system to demonstrate that profilin, a host actin monomer-binding protein, is necessary for bacterial actin-based motility. We find that extracts from which profilin has been depleted do not support comet tail formation or bacterial motility. In extracts and host cells, profilin is localized to the back half of the surface of motile L. monocytogenes, the site of actin filament assembly in the tail. This association is not observed with L. monocytogenes mutants that do not express the ActA protein, a bacterial gene product necessary for motility and virulence. Profilin also fails to bind L. monocytogenes grown outside of host cytoplasm, suggesting that at least one other host cell factor is required for this association.
AB - Within hours of Listeria monocytogenes infection, host cell actin filaments form a dense cloud around the intracytoplasmic bacteria and then rearrange to form a polarized comet tail that is associated with moving bacteria. We have devised a cell-free extract system capable of faithfully reconstituting L. monocytogenes motility, and we have used this system to demonstrate that profilin, a host actin monomer-binding protein, is necessary for bacterial actin-based motility. We find that extracts from which profilin has been depleted do not support comet tail formation or bacterial motility. In extracts and host cells, profilin is localized to the back half of the surface of motile L. monocytogenes, the site of actin filament assembly in the tail. This association is not observed with L. monocytogenes mutants that do not express the ActA protein, a bacterial gene product necessary for motility and virulence. Profilin also fails to bind L. monocytogenes grown outside of host cytoplasm, suggesting that at least one other host cell factor is required for this association.
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U2 - 10.1016/0092-8674(94)90114-7
DO - 10.1016/0092-8674(94)90114-7
M3 - Article
C2 - 8313471
AN - SCOPUS:0028173688
VL - 76
SP - 505
EP - 517
JO - Cell
JF - Cell
SN - 0092-8674
IS - 3
ER -