University of Massachusetts Medical School Faculty Publications
UMMS Affiliation
RNA Therapeutics Institute
Publication Date
2018-04-11
Document Type
Article Preprint
Disciplines
Genetic Phenomena | Molecular Biology
Abstract
Single-molecule binding assays enable the study of how molecular machines assemble and function. Current algorithms can identify and locate individual molecules, but require tedious manual validation of each spot. Moreover, no solution for high-throughput analysis of single-molecule binding data exists. Here, we describe an automated pipeline to analyze single-molecule data over a wide range of experimental conditions. We benchmarked the pipeline by measuring the binding properties of the well-studied, DNA-guided DNA endonuclease, TtAgo, an Argonaute protein from the Eubacterium Thermus thermophilus. We also used the pipeline to extend our understanding of TtAgo by measuring the protein's binding kinetics at physiological temperatures and for target DNAs containing multiple, adjacent binding sites.
Keywords
Automated Bayesian Pipeline, Single-Molecule Binding Data, TtAgo, Thermus thermophilus, DNA, Molecular Biology
Rights and Permissions
The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It is made available under a CC-BY-NC-ND 4.0 International license.
DOI of Published Version
10.1101/261917
Source
bioRxiv 261917; doi: https://doi.org/10.1101/26191. Link to preprint on bioRxiv service.
Journal/Book/Conference Title
bioRxiv
Repository Citation
Smith, Carlas; Jouravleva, Karina; Huisman, Maximiliaan; Jolly, Samson M.; Zamore, Phillip D.; and Grünwald, David, "An Automated Bayesian Pipeline for Rapid Analysis of Single-Molecule Binding Data" (2018). University of Massachusetts Medical School Faculty Publications. 1499.
https://escholarship.umassmed.edu/faculty_pubs/1499
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.