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PMID: 9885246 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Coronin promotes the rapid assembly and cross-linking of actin filaments and may link the actin and microtubule cytoskeletons in yeast.

The Journal of cell biology ·Vol. 144 ·No. 1 ·1999-01-11 ·Pages 83-98

Goode BL, Wong JJ, Butty AC, Peter M, McCormack AL, Yates JR, Drubin DG, Barnes G

Abstract

Coronin is a highly conserved actin-associated protein that until now has had unknown biochemical activities. Using microtubule affinity chromatography, we coisolated actin and a homologue of coronin, Crn1p, from Saccharomyces cerevisiae cell extracts. Crn1p is an abundant component of the cortical actin cytoskeleton and binds to F-actin with high affinity (Kd 6 x 10(-9) M). Crn1p promotes the rapid barbed-end assembly of actin filaments and cross-links filaments into bundles and more complex networks, but does not stabilize them. Genetic analyses with a crn1Delta deletion mutation also are consistent with Crn1p regulating filament assembly rather than stability. Filament cross-linking depends on the coiled coil domain of Crn1p, suggesting a requirement for Crn1p dimerization. Assembly-promoting activity is independent of cross-linking and could be due to nucleation and/or accelerated polymerization. Crn1p also binds to microtubules in vitro, and microtubule binding is enhanced by the presence of actin filaments. Microtubule binding is mediated by a region of Crn1p that contains sequences (not found in other coronins) homologous to the microtubule binding region of MAP1B. These activities, considered with microtubule defects observed in crn1Delta cells and in cells overexpressing Crn1p, suggest that Crn1p may provide a functional link between the actin and microtubule cytoskeletons in yeast.

MeSH Terms
Actin Cytoskeleton/metabolism Actins/isolation & purification,metabolism Amino Acid Sequence Animals Binding Sites Cell Division Chromatography, Affinity Cross-Linking Reagents Cytoskeleton/metabolism Fungal Proteins/genetics,metabolism Genes, Fungal Glutathione Transferase/genetics,metabolism Mice Microfilament Proteins/genetics,metabolism Microtubules/metabolism Molecular Sequence Data Mutagenesis Recombinant Fusion Proteins/genetics,metabolism Saccharomyces cerevisiae/genetics,metabolism
Chemicals
Actins Cross-Linking Reagents Fungal Proteins Microfilament Proteins Recombinant Fusion Proteins coronin proteins Glutathione Transferase
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Goode B L
Department of Molecular and Cell Biology, University of California, Berkeley, California 94720-3202, USA.
Wong J J
Butty A C
Peter M
McCormack A L
Yates J R
Drubin D G
Barnes G
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.144.1.83
Published
1999-01-11
Pages
83-98
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2148128
Subset
IM
Grants
NIGMS NIH HHS · GM42759 · United States
NIGMS NIH HHS · GM17715-02 · United States
NIGMS NIH HHS · GM47842 · United States
External Links
PubMed source DOI: 10.1083/jcb.144.1.83 PMC Full Text View journal details
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