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Interaction of the Proline-Rich Domain of Fission Yeast WASp (Ws1p1) with Actin Filaments

Aaron D. Rosenbloom, View ORCID ProfileThomas D. Pollard
doi: https://doi.org/10.1101/2022.01.26.477833
Aaron D. Rosenbloom
1Department of Chemistry, Yale University, PO Box 208107, New Haven, CT 06520-8103 USA
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Thomas D. Pollard
2Departments of Molecular Cellular and Developmental Biology, of Molecular Biophysics and Biochemistry and of Cell Biology, Yale University, PO Box 208103, New Haven, CT 06520-8103 USA
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  • ORCID record for Thomas D. Pollard
  • For correspondence: thomas.pollard@yale.edu
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Abstract

Background The Wiskott-Aldrich Syndrome protein (WASp) family of proteins plays a crucial role in the activation of the Arp2/3 (actin-related protein 2/3) complex to promote the branching of actin filaments. The proline-rich domain (PRD) of WASp is known to contribute to branching nucleation but was overlooked, until experiments showed that the PRD of budding yeast Las17 can bind actin filaments (1).

Methods We purified recombinant proline-rich domains from fission yeast S. pombe Wsp1 and budding yeast S. cerevisiae Las17 to test in biochemical assays of actin binding and polymerization.

Results The PRD of the S. pombe Wsp1 binds actin filaments with micromolar affinity. The PRDs of both Wsp1 and Las17 slowed the rate of actin filament elongation by Mg-ATP-actin monomers by half and slowed the spontaneous polymerization of Mg-ATP-actin monomers modestly.

Conclusion The affinity of PRDs of WASp-family proteins for actin filaments is high enough to contribute to the reported stimulation of actin filament branching by Arp2/3 complex.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY 4.0 International license.
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Posted January 26, 2022.
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Interaction of the Proline-Rich Domain of Fission Yeast WASp (Ws1p1) with Actin Filaments
Aaron D. Rosenbloom, Thomas D. Pollard
bioRxiv 2022.01.26.477833; doi: https://doi.org/10.1101/2022.01.26.477833
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Interaction of the Proline-Rich Domain of Fission Yeast WASp (Ws1p1) with Actin Filaments
Aaron D. Rosenbloom, Thomas D. Pollard
bioRxiv 2022.01.26.477833; doi: https://doi.org/10.1101/2022.01.26.477833

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