title: Towards physiological complexity with in vitro single-molecule biophysics.
abstract: Single-molecule biology has matured in recent years, driven to greater sophistication by the development of increasingly advanced experimental techniques. A progressive appreciation for its unique strengths is attracting research that spans an exceptionally broad swath of physiological phenomena--from the function of nucleosomes to protein diffusion in the cell membrane. Newfound enthusiasm notwithstanding, the single-molecule approach is limited to an intrinsically defined set of biological questions; such limitation applies to all experimental approaches, and an explicit statement of the boundaries delineating each set offers a guide to most fruitfully orienting in vitro single-molecule research in the future. Here, we briefly describe a simple conceptual framework to categorize how submolecular, molecular and intracellular processes are studied. We highlight the domain of single-molecule biology in this scheme, with an emphasis on its ability to probe various forms of heterogeneity inherent to populations of discrete biological macromolecules. We then give a general overview of our high-throughput DNA curtain methodology for studying protein-nucleic acid interactions, and by contextualizing it within this framework, we explore what might be the most enticing avenues of future research. We anticipate that a focus on single-molecule biology's unique strengths will suggest a new generation of experiments with greater complexity and more immediately translatable physiological relevance.
authors.affiliation: Department of Biological Sciences, Columbia University, 650 West 168th Street, New York, NY 10032, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Duzdevich
authors.forename: Eric C
authors.initials: EC
authors.lastname: Greene
mesh_terms: D000818:Animals; D001692:Biological Transport; D055592:Biophysical Phenomena; D002462:Cell Membrane / Q000737:chemistry; D058948:Cell Tracking / Q000379:methods; D042002:Chromatin Assembly and Disassembly; D004247:DNA / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D056890:Eukaryota / Q000737:chemistry / Q000166:cytology*; D005456:Fluorescent Dyes / Q000737:chemistry; D059014:High-Throughput Nucleotide Sequencing; D007091:Image Processing, Computer-Assisted / Q000379:methods; D008051:Lipid Bilayers / Q000737:chemistry; D008856:Microscopy, Fluorescence; D009707:Nucleosomes / Q000737:chemistry*; D025941:Protein Interaction Mapping
chemical_list: D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D009707:Nucleosomes; D004247:DNA