Engineering allostery

Raman, Srivatsan; Taylor, Noah; Genuth, Naomi; Fields, Stanley; Church, George M.

doi:10.1016/j.tig.2014.09.004

Title: Engineering allostery

Journal Article · Wed Oct 08 00:00:00 EDT 2014 · Trends in Genetics

DOI:https://doi.org/10.1016/j.tig.2014.09.004· OSTI ID:1344516

Raman, Srivatsan ^[1]; Taylor, Noah ^[1]; Genuth, Naomi ^[1]; Fields, Stanley ^[2]; Church, George M. ^[1]

Harvard Univ., Boston, MA (United States); Harvard Medical School, Boston, MA (United States)
Univ. of Washington, Seattle, WA (United States)

Allosteric proteins have great potential in synthetic biology, but our limited understanding of the molecular underpinnings of allostery has hindered the development of designer molecules, including transcription factors with new DNA-binding or ligand-binding specificities that respond appropriately to inducers. Such allosteric proteins could function as novel switches in complex circuits, metabolite sensors, or as orthogonal regulators for independent, inducible control of multiple genes. Advances in DNA synthesis and next-generation sequencing technologies have enabled the assessment of millions of mutants in a single experiment, providing new opportunities to study allostery. Using the classic LacI protein as an example, in this paper we describe a genetic selection system using a bidirectional reporter to capture mutants in both allosteric states, allowing the positions most crucial for allostery to be identified. Finally, this approach is not limited to bacterial transcription factors, and could reveal new mechanistic insights and facilitate engineering of other major classes of allosteric proteins such as nuclear receptors, two-component systems, G protein-coupled receptors, and protein kinases.

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Research Organization:: Harvard Univ., Cambridge, MA (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: FG02-02ER63445

OSTI ID:: 1344516

Alternate ID(s):: OSTI ID: 1227554

Journal Information:: Trends in Genetics, Vol. 30, Issue 12; ISSN 0168-9525

Publisher:: ElsevierCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 47 works

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