Common formin-regulating sequences in Smy1 and Bud14 are required for the control of actin cable assembly in vivo

Molecular Biology of the Cell
Julian A EskinBruce L Goode

Abstract

Formins comprise a large family of proteins with diverse roles in remodeling the actin cytoskeleton. However, the spatiotemporal mechanisms used by cells to control formin activities are only beginning to be understood. Here we dissected Smy1, which has dual roles in regulating formins and myosin. Using mutagenesis, we identified specific sequences in Smy1 critical for its in vitro inhibitory effects on the FH2 domain of the formin Bnr1. By integrating smy1 alleles targeting those sequences, we genetically uncoupled Smy1's functions in regulating formins and myosin. Quantitative imaging analysis further demonstrated that the ability of Smy1 to directly control Bnr1 activity is crucial in vivo for proper actin cable length, shape, and velocity and, in turn, efficient secretory vesicle transport. A Smy1-like sequence motif was also identified in a different Bnr1 regulator, Bud14, and found to be essential for Bud14 functions in regulating actin cable architecture and function in vivo. Together these observations reveal unanticipated mechanistic ties between two distinct formin regulators. Further, they emphasize the importance of tightly controlling formin activities in vivo to generate specialized geometries and dynamics of acti...Continue Reading

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Citations

May 21, 2016·Science·Jessica L Henty-RidillaBruce L Goode
May 21, 2016·Science·Klemens Rottner
Jun 17, 2016·Molecular Biology of the Cell·Kyaw Myo LwinAnthony Bretscher
Apr 18, 2016·Cellular and Molecular Life Sciences : CMLS·Maria Angeles Juanes, Simonetta Piatti
Feb 26, 2019·The Journal of Biological Chemistry·Kun TangZhiyi Wei
Aug 5, 2018·The Journal of Cell Biology·Mikael V GarabedianBruce L Goode
Jun 25, 2020·Molecular Biology of the Cell·Mikael V GarabedianBruce L Goode
Feb 27, 2021·Frontiers in Cell and Developmental Biology·Thomas J Rands, Bruce L Goode

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Methods Mentioned

BETA
Dissection
restriction digest
fluorescence microscopy

Software Mentioned

MATLAB
NIS Elements Advanced Research
BLAST
OpenLab
ImageJ

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