Student or educator? Sign up with your school email and get 6 months of Pro free.
CoChat
Sign Up
Cited by 1Gene Regulatory Network AnalysisBacterial Genetics and BiotechnologyMicrobial Metabolic Engineering and Bioproduction

The DOI matches its Crossref record; 55 of the 55 references it cites were found in a registry or an index.

Fabrication, Evolution, and Mutual Conversion of d -Fucose-Activatable and -Repressible Acetyltransferase upon Mutations

Open access
Yanai, Yuki, Tsukada, Miyu, Kimura, Yuki, Umeno, Daisuke·2025·ACS Synthetic Biology
DOI 10.1021/acssynbio.4c00169

Published version of a preprint

Open this paper in CoChat — free account

Abstract

Abstract The fusion of different proteins can result in the linkage-dependent emergence of molecular switches. In cases where allosteric regulation is designed between the input and output modules of fusion proteins, it is hard to predict whether on-switching or off-switching will occur. However, binding-induced folding, a non-allosteric molecular switch mechanism, has the potential to quickly establish a mutually regulatory relationship between the two fused proteins, in the way whether on-switching or off-switching will occur would be predictable. We inserted chloramphenicol acetyltransferase (CAT) from E. coli into a loop of a D-fucose-responsive mutant of transcription factor AraC, using linker libraries with various lengths. We found that on-switches tend to emerge when two proteins are fused with a small pitch gap at the junction, while fusion designs with a large pitch gap result in the frequent emergence of off-switches. Both types of switches rapidly evolved their switching efficiency upon mutations, establishing the D-fucose-on and -off regulation of CAT activity without disrupting the D-fucose-inducible logic of AraC function. To our surprise, both one-input/two-output split gates thus obtained could be easily inter-converted upon mutations. Through mutations, proteins not only frequently acquire properties as binding-induced folders, but also rapidly establish and evolve a mutual regulatory relationship with unrelated fusion partners, as well as transform their regulatory logic.

Overview

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1021/acssynbio.4c00169

Cites

55 of 55 references verified0 citation errors · 0 not checkable

Statistics

Verification

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Citations

Cites

Research works printed but not linked: 2

  1. Liu Z., 2023, bioRxiv
  2. 10.1111/j.1574-6976.2010.00226.x · DOI

Metadata

Authors
Yanai, Y., Tsukada, M., Kimura, Y., & Umeno, D.
Affiliations
  • Waseda University (JP)
  • Chiba University (JP)
Corresponding author
Umeno, Daisuke
ORCID
Venue
ACS Synthetic Biology
Publisher
American Chemical Society (ACS)
Published
2025-04-30
Volume
14
Issue
6
Pages
1936-1947
ISSN
2161-5063 (ISSN-L)
DOI
10.1021/acssynbio.4c00169
OpenAlex
W4409983053
Cited by
1
References
55
Citations by year
2026: 1
FWCI
0.30
Type
journal article
Language
en
Open access
Licence
Locations
  • ACS Synthetic Biology (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Funders
  • New Energy and Industrial Technology Development Organization (JPNP20011)
  • Japan Society for the Promotion of Science (16H06450, 18H01791, 21H01721)
  • KIOXIA Holdings Corporation
  • Futaba Electronics Memorial Foundation
  • Salt Science Research Foundation (2201)
  • Waseda University (22P04)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Gene Regulatory Network Analysis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Keywords
AraC, acetyltransferase, split gate, on-off switch, linker length, protein fusion, random mutagenesis, protein evolution, mutual regulation, transcriptional regulation
MeSH terms
Escherichia coli, Fucose, Mutation, Recombinant Fusion Proteins, Chloramphenicol O-Acetyltransferase, Escherichia coli Proteins

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review

Data sources

Bibliographic data: Semantic Scholar, OpenAlex, Crossref.

Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).

Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.