Cytoskeletal disassembly by optogenetic control of RhoA signaling termination
- 1University of Pennsylvania
Abstract
Cellular morphodynamics require adaptive cytoskeletal remodeling, mediated by precisely coordinated activation and termination of RhoA GTPase signaling. RhoA activation is well-studied, but the kinetics and molecular basis of signaling termination remain poorly understood. We engineered an optogenetic toolbox on the single-component BcLOV4 platform for bidirectional control of RhoA activation (opto-GEF11) and termination (opto-DLC1). Prior studies have inferred GTPase inactivation kinetics indirectly, by tracking passive recovery from an activated state, but whether this reflects the kinetics of forward signaling termination remains unclear. Using opto-DLC1, we show that direct RhoA termination is an order of magnitude faster than passive disactivation, despite comparable signaling amplitude. Mechanistically, opto-DLC1 triggered rapid actin disassembly through cofilin disinhibition but drove YAP nuclear efflux at half the rate of opto-GEF11-induced nuclear influx. Together, these findings introduce a platform technology for controlling protein signaling termination, resolve RhoA activation and termination kinetics with sub-second precision, and reveal a mechanistic asymmetry between signaling activation and termination that enables cytoskeletal homeostasis.
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BibTeX
@misc{berlew2026cytoskeletal,
title = {{Cytoskeletal disassembly by optogenetic control of RhoA signaling termination}},
author = {E. E. Berlew and J. Barakat and P. Camacho Sierra and J. D. Boerckel},
howpublished = {bioRxiv},
year = {2026},
month = sep,
doi = {10.64898/2026.09.03.749286},
url = {https://www.biorxiv.org/content/10.64898/2026.09.03.749286v1},
note = {CC BY-NC},
}
RIS
TY - UNPB
TI - Cytoskeletal disassembly by optogenetic control of RhoA signaling termination
AU - E. E. Berlew
AU - J. Barakat
AU - P. Camacho Sierra
AU - J. D. Boerckel
PY - 2026
DA - 2026/09/03
PB - bioRxiv
DO - 10.64898/2026.09.03.749286
UR - https://www.biorxiv.org/content/10.64898/2026.09.03.749286v1
AB - Cellular morphodynamics require adaptive cytoskeletal remodeling, mediated by precisely coordinated activation and termination of RhoA GTPase signaling. RhoA activation is well-studied, but the kinetics and molecular basis of signaling termination remain poorly understood. We engineered an optogenetic toolbox on the single-component BcLOV4 platform for bidirectional control of RhoA activation (opto-GEF11) and termination (opto-DLC1). Prior studies have inferred GTPase inactivation kinetics indirectly, by tracking passive recovery from an activated state, but whether this reflects the kinetics of forward signaling termination remains unclear. Using opto-DLC1, we show that direct RhoA termination is an order of magnitude faster than passive disactivation, despite comparable signaling amplitude. Mechanistically, opto-DLC1 triggered rapid actin disassembly through cofilin disinhibition but drove YAP nuclear efflux at half the rate of opto-GEF11-induced nuclear influx. Together, these findings introduce a platform technology for controlling protein signaling termination, resolve RhoA activation and termination kinetics with sub-second precision, and reveal a mechanistic asymmetry between signaling activation and termination that enables cytoskeletal homeostasis.
ER -