DNA sequences for: Synthetic control of actin polymerization and symmetry breaking in active protocells
Data files
May 20, 2024 version files 65.23 KB
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2xstrep-mActA(1-184)-FRB-CFP(pET100).gbk
17.15 KB
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Cap_32(pET14b).gbk
2.99 KB
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Cap_34(pET14b).gbk
2.36 KB
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CFP-FRB(pET28a).gbk
4.98 KB
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mActA(1-584)-FKBP-CFP(pET28a).gbk
7.08 KB
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mCherry-FKBP-MARCKS(pBiEx1).gbk
10.98 KB
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mCherry-FKBP(pBiEx1).gbk
10.56 KB
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README.md
1.90 KB
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YFP-FKBP_full_length_(EYFP-C1).gbk
7.23 KB
Abstract
Non-linear biomolecular interactions on membranes drive membrane remodeling crucial for biological processes including chemotaxis, cytokinesis, and endocytosis. The complexity of biomolecular interactions, their redundancy, and the importance of spatiotemporal context in membrane organization impede understanding of the physical principles governing membrane mechanics. Developing a minimal in vitro system that mimics molecular signaling and mem- brane remodeling while maintaining physiological fidelity poses a significant challenge. Inspired by chemotaxis, we reconstructed chemically regulated actin polymerization inside vesicles, guiding membrane self-organization. An external, undirected chemical input induced directed actin polymerization and membrane deformation uncorrelated with upstream biochemical cues, suggesting symmetry breaking. A biophysical model incorporating actin dynamics and membrane mechanics proposes that uneven actin distributions cause non-linear membrane deformations, consistent with experimental findings. This protocellular system illuminates the interplay between actin dynamics and membrane shape during symmetry breaking, offering insights into chemotaxis and other cell biological processes.
Supplementary DNA Sequences for ‘Synthetic Control of Actin Polymerization and Symmetry Breaking in Active Protocells’
Manuscript Title: Synthetic Control of Actin Polymerization and Symmetry Breaking in Active Protocells
Authors: Shiva Razavi, Felix Wong, Bedri Abubaker-Sharif, Hideaki T. Matsubayashi, Hideki Nakamura, Nhung Thi Hong Nguyen, Douglas N. Robinson, Baoyu Chen, Pablo A. Iglesias, and Takanari Inoue.
Access this dataset on Dryad: https://doi.org/10.5061/dryad.rxwdbrvh9
Description of the data and file structure
This dataset comprises the DNA sequences corresponding to the proteins that were purified and utilized in this study.
The files are provided in Genebank (.gbk) format and include the following sequences:
- 2xStrep-mActA(1-184)-FRB-CFP: Contains the sequence of ActA(1-184) codon-optimized for mammalian expression and tagged with a Strep tag for purification. The FRB module enables the translocation of ActA via chemically inducible dimerization. The CFP tag facilitates visualization using standard microscopy techniques.
- mActA(1-584)-FKBP-CFP: Contains the sequence of ActA(1-584) codon-optimized for mammalian expression and tagged with a 6xHis tag for purification.
- Cap_32 and Cap_34: These are the dictyostelium capping protein sequences used in this study.
- YFP-FKBP and CFP-FRB: Contain chemically inducible dimerization modules that dimerize with rapamycin, resulting in a FRET signal.
- mCherry-FKBP-MARCKS: Contains FKBP tagged with MARCKS for lipid tethering.
- mCherry-FKBP: Contains FKBP tagged with a 6xHis tag for tethering to nickel-conjugated lipid.
Sharing/Access information
To received a vial of the DNA sequences please contact the corresponding authors: srazavi@mit.edu and jctinoue@jhmi.edu
Sequencing Files: The sequences provided were generated using standard cloning methods and were verified via Sanger sequencing for accuracy and completeness.