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PMID: 8909540 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

The tandem repeat domain in the Listeria monocytogenes ActA protein controls the rate of actin-based motility, the percentage of moving bacteria, and the localization of vasodilator-stimulated phosphoprotein and profilin.

The Journal of cell biology ·Vol. 135 ·No. 3 ·1996-11-00 ·Pages 647-60

Smith GA, Theriot JA, Portnoy DA

Abstract

The ActA protein is responsible for the actin-based movement of Listeria monocytogenes in the cytosol of eukaryotic cells. Analysis of mutants in which we varied the number of proline-rich repeats (PRR; consensus sequence DFPPPPTDEEL) revealed a linear relationship between the number of PRRs and the rate of movement, with each repeat contributing approximately 2-3 microns/min. Mutants lacking all functional PRRs (generated by deletion or point mutation) moved at rates 30% of wild-type. Indirect immunofluorescence indicated that the PRRs were directly responsible for binding of vasodilator-stimulated phosphoprotein (VASP) and for the localization of profilin at the bacterial surface. The long repeats, which are interdigitated between the PRRs, increased the frequency with which actin-based motility occurred by a mechanism independent of the PRRs, VASP, and profilin. Lastly, a mutant which expressed low levels of ActA exhibited a phenotype indicative of a threshold; there was a very low percentage of moving bacteria, but when movement did occur, it was at wild-type rates. These results indicate that the ActA protein directs at least three separable events: (1) initiation of actin polymerization that is independent of the repeat region; (2) initiation of movement dependent on the long repeats and the amount of ActA; and (3) movement rate dependent on the PRRs.

MeSH Terms
Actins/analysis,biosynthesis Animals Bacterial Proteins/analysis,genetics,physiology Cell Adhesion Molecules/analysis Cell Line Contractile Proteins/analysis DNA, Bacterial/genetics Humans Lethal Dose 50 Listeria monocytogenes/chemistry,genetics,pathogenicity,physiology Membrane Proteins/genetics,physiology Mice Mice, Inbred BALB C Microfilament Proteins/analysis Mutation Phosphoproteins/analysis Polymers Profilins Proline Repetitive Sequences, Nucleic Acid/genetics
Chemicals
Actins Bacterial Proteins Cell Adhesion Molecules Contractile Proteins DNA, Bacterial Membrane Proteins Microfilament Proteins Pfn1 protein, mouse Phosphoproteins Polymers Profilins vasodilator-stimulated phosphoprotein actA protein, Listeria monocytogenes Proline
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Smith G A
Department of Microbiology, University of Pennsylvania, School of Medicine, Philadelphia 19104-4318, USA.
Theriot J A
Portnoy D A
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.135.3.647
Published
1996-11-00
Pages
647-60
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2121076
Subset
IM
Grants
NIAID NIH HHS · R37 AI036929 · United States
NIAID NIH HHS · AI-26917 · United States
NIAID NIH HHS · AI-36827 · United States
External Links
PubMed source DOI: 10.1083/jcb.135.3.647 PMC Full Text View journal details
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