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PMID: 15385960 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Mutant actins that stabilise F-actin use distinct mechanisms to activate the SRF coactivator MAL.

The EMBO journal ·Vol. 23 ·No. 20 ·2004-10-13 ·Pages 3973-83

Posern G, Miralles F, Guettler S, Treisman R

Abstract

Nuclear accumulation of the serum response factor coactivator MAL/MKL1 is controlled by its interaction with G-actin, which results in its retention in the cytoplasm in cells with low Rho activity. We previously identified actin mutants whose expression promotes MAL nuclear accumulation via an unknown mechanism. Here, we show that actin interacts directly with MAL in vitro with high affinity. We identify a further activating mutation, G15S, which stabilises F-actin, as do the activating actins S14C and V159N. The three mutants share several biochemical properties, but can be distinguished by their ability to bind cofilin, ATP and MAL. MAL interaction with actin S14C is essentially undetectable, and that with actin V159N is weakened. In contrast, actin G15S interacts more strongly with MAL than the wild-type protein. Strikingly, the nuclear accumulation of MAL induced by overexpression of actin S14C is substantially dependent on Rho activity and actin treadmilling, while that induced by actin G15S expression is not. We propose a model in which actin G15S acts directly to promote MAL nuclear entry.

MeSH Terms
Actin Depolymerizing Factors Actins/analysis,genetics,metabolism Active Transport, Cell Nucleus/genetics Adenosine Triphosphate/metabolism Amino Acid Motifs/genetics Amino Acid Sequence Animals Binding Sites Cell Extracts Cell Nucleus/metabolism DNA-Binding Proteins/chemistry,genetics,metabolism Deoxyribonuclease I/metabolism Genes, Reporter Glutathione Transferase/metabolism Luciferases/metabolism Mice Microfilament Proteins/metabolism NIH 3T3 Cells Oncogene Proteins, Fusion/chemistry,genetics,metabolism Phalloidine/metabolism Point Mutation Precipitin Tests Protein Binding Serum Response Factor/metabolism Signal Transduction Trans-Activators Two-Hybrid System Techniques
Chemicals
Actin Depolymerizing Factors Actins Cell Extracts DNA-Binding Proteins MRTFA protein, human Microfilament Proteins Oncogene Proteins, Fusion Serum Response Factor Trans-Activators Phalloidine Adenosine Triphosphate Luciferases Glutathione Transferase Deoxyribonuclease I
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Posern Guido
Transcription Laboratory, Cancer Research UK London Research Institute, Lincoln's Inn Fields Laboratories, London, UK.
Miralles Francesc
Guettler Sebastian
Treisman Richard
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1038/sj.emboj.7600404
Published
2004-10-13
Epub
2004-00-23
Pages
3973-83
Language
English
Region
England
NLM ID
8208664
PMCID
PMC524340
Subset
IM
External Links
PubMed source DOI: 10.1038/sj.emboj.7600404 PMC Full Text View journal details
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