@inbook{30e164883dc34a279cb168741806106d,
title = "A Fluorescent Assay to Monitor Ligand-Dependent Closure of the Hexameric Rho Helicase Ring",
abstract = "The bacterial Rho protein is an exemplar RecA-family hexameric helicase that assists with the termination of RNA polymerase activity on a variety of transcripts. During its catalytic cycle, Rho both loads onto and translocates along RNA through a series of tightly regulated, ligand-dependent conformational changes. Here we describe an assay to track Rho as it switches from an open-ring (RNA-loading) to a closed-ring (RNA-translocation) configuration by monitoring the association of a fluorescein-labeled RNA to Rho{\textquoteright}s central pore as a change in fluorescence anisotropy. The assay, which is in principle adaptable to the study of ligand-dependent isomerization events in other ring-shaped translocases, is readily amenable to 384-well format plates and small-molecule screening efforts.",
keywords = "ATPase, Fluorescence anisotropy, Hexameric helicase, High-throughput screening, RNA, Rho, Ring translocase, Transcription termination",
author = "Lawson, {Michael R.} and Berger, {James M.}",
note = "Funding Information: This work was supported by G. Harold and Leila Y. Mathers Foundation and the National Institute of General Medical Sciences (R37-071747), to J.M.B. M.R.L. gratefully acknowledges support from the A.P. Giannini Foundation. Publisher Copyright: {\textcopyright} 2021, Springer Science+Business Media, LLC, part of Springer Nature.",
year = "2021",
doi = "10.1007/978-1-0716-0935-4_9",
language = "English (US)",
series = "Methods in Molecular Biology",
publisher = "Humana Press Inc.",
pages = "133--142",
booktitle = "Methods in Molecular Biology",
}