---
DOAJ_listed: '1'
OA_place: publisher
OA_type: gold
PlanS_conform: '1'
_id: '21928'
abstract:
- lang: eng
text: Antibiotic combination in time and space is a key strategy to combat antimicrobial
resistance. The success of such treatment designs requires their robust efficacy
across treatment conditions and a pathogen’s genomic diversity. This study found
that an initial treatment with a β-lactam antibiotic causes robust cellular sensitization
towards an aminoglycoside antibiotic across the high-risk human pathogen Pseudomonas
aeruginosa, including resistant strains. This phenomenon of cellular sensitization,
termed negative hysteresis, is modulated by the Cpx envelope stress response system
and linked to membrane stress during growth. The increase in efficacy is achieved
through a β-lactam induced elevated cellular uptake of the subsequently administered
aminoglycoside. Negative hysteresis and the Cpx system are linked in several cases
to the expression of synergistic drug interactions, thus enhancing efficacy of
antibiotic combinations. Overall, our study identifies the phenomenon of negative
hysteresis as a robustly inducible phenotype and thus a unique focus for optimizing
antimicrobial therapy.
acknowledgement: "We are very grateful to S. Hernando-Amado (Madrid, Spain), C. Pál
(Szeged, Hungary), and T. Bollenbach (Cologne, Germany) for critical comments and
advice on the manuscript. We further thank D. Rogers, J. Summers (Ploen, Germany)
for guidance in allelic exchange, P. Rainey (Ploen, Germany) for providing the plasmids
and strains, then J. Lorenzen, K. Flinder, N. Steinbach, S. Butze (all Schulenburg
lab), and L. Kirchhoff (Rupp lab) for supporting the experimental work, and also
the Rupp and Schulenburg groups for general feedback. We are grateful for financial
support from the German Research Foundation within the Research and Training Group
2501 (RTG 2501) on Translational Evolutionary Research (project 4.2 to H.S.), within
the Excellence cluster Precision Medicine in chronic Inflammation (PMI; funding
under Germany’s Excellence Strategy EXC 2167-390884018, to B.K., K.R., J.R., H.S.),
within the Clinician Scientist Program in Evolutionary Medicine (CSEM) – project
number 413490537 (to EEG), and as part of the individual grants SCHU 1415/12-2 (to
H.S.) and BR-2915/7-1 (to M.B.). We are grateful for financial support from the
Swedish Research Council, project number 2021-02091 (to D.I.A.). We are also grateful
for financial support from the Max-Planck Society (Fellowship to H.S.), the Leibniz
Association within the Leibniz Science-Campus Evolutionary Medicine of the Lung
(EvoLUNG, to H.S.), and the project SKILLED funded by the DAMP foundation (to J.R.,
H.S.). This work was also supported by the ZMB Young Scientist award and the FWF
grant 10.55776/ESP219 (to R.R.) and the TransEvo Innovation prize (to F.B.). The
funders had no role in study design, data collection and interpretation, or the
decision to submit the work for publication. Open Access funding enabled and organized
by Projekt DEAL.\r\n"
article_number: '4487'
article_processing_charge: Yes (via OA deal)
article_type: original
author:
- first_name: Florian
full_name: Buchholz, Florian
last_name: Buchholz
- first_name: Lina M.
full_name: Upterworth, Lina M.
last_name: Upterworth
- first_name: Leif
full_name: Tueffers, Leif
last_name: Tueffers
- first_name: Espen E.
full_name: Groth, Espen E.
last_name: Groth
- first_name: Kira
full_name: Haas, Kira
last_name: Haas
- first_name: Daniel
full_name: Schütz, Daniel
last_name: Schütz
- first_name: Abigail
full_name: Savietto Scholz, Abigail
last_name: Savietto Scholz
- first_name: Aditi
full_name: Batra, Aditi
last_name: Batra
- first_name: Surajit
full_name: Pal, Surajit
last_name: Pal
- first_name: Samarpita
full_name: Banerjee, Samarpita
last_name: Banerjee
- first_name: Badri N.
full_name: Dubey, Badri N.
last_name: Dubey
- first_name: Sören
full_name: Franzenburg, Sören
last_name: Franzenburg
- first_name: Barbara
full_name: Kalsdorf, Barbara
last_name: Kalsdorf
- first_name: Klaus F.
full_name: Rabe, Klaus F.
last_name: Rabe
- first_name: Dennis
full_name: Nurjadi, Dennis
last_name: Nurjadi
- first_name: Jan
full_name: Rupp, Jan
last_name: Rupp
- first_name: Dan I.
full_name: Andersson, Dan I.
last_name: Andersson
- first_name: Holger
full_name: Sondermann, Holger
last_name: Sondermann
- first_name: Marc
full_name: Bramkamp, Marc
last_name: Bramkamp
- first_name: Roderich
full_name: Römhild, Roderich
id: 68E56E44-62B0-11EA-B963-444F3DDC885E
last_name: Römhild
orcid: 0000-0001-9480-5261
- first_name: Hinrich
full_name: Schulenburg, Hinrich
last_name: Schulenburg
citation:
ama: Buchholz F, Upterworth LM, Tueffers L, et al. Robust antibiotic sensitization
of pathogenic Pseudomonas aeruginosa via negative hysteresis in the cell envelope.
Nature Communications. 2026;17. doi:10.1038/s41467-026-71178-5
apa: Buchholz, F., Upterworth, L. M., Tueffers, L., Groth, E. E., Haas, K., Schütz,
D., … Schulenburg, H. (2026). Robust antibiotic sensitization of pathogenic Pseudomonas
aeruginosa via negative hysteresis in the cell envelope. Nature Communications.
Springer Nature. https://doi.org/10.1038/s41467-026-71178-5
chicago: Buchholz, Florian, Lina M. Upterworth, Leif Tueffers, Espen E. Groth, Kira
Haas, Daniel Schütz, Abigail Savietto Scholz, et al. “Robust Antibiotic Sensitization
of Pathogenic Pseudomonas Aeruginosa via Negative Hysteresis in the Cell Envelope.”
Nature Communications. Springer Nature, 2026. https://doi.org/10.1038/s41467-026-71178-5.
ieee: F. Buchholz et al., “Robust antibiotic sensitization of pathogenic
Pseudomonas aeruginosa via negative hysteresis in the cell envelope,” Nature
Communications, vol. 17. Springer Nature, 2026.
ista: Buchholz F, Upterworth LM, Tueffers L, Groth EE, Haas K, Schütz D, Savietto
Scholz A, Batra A, Pal S, Banerjee S, Dubey BN, Franzenburg S, Kalsdorf B, Rabe
KF, Nurjadi D, Rupp J, Andersson DI, Sondermann H, Bramkamp M, Römhild R, Schulenburg
H. 2026. Robust antibiotic sensitization of pathogenic Pseudomonas aeruginosa
via negative hysteresis in the cell envelope. Nature Communications. 17, 4487.
mla: Buchholz, Florian, et al. “Robust Antibiotic Sensitization of Pathogenic Pseudomonas
Aeruginosa via Negative Hysteresis in the Cell Envelope.” Nature Communications,
vol. 17, 4487, Springer Nature, 2026, doi:10.1038/s41467-026-71178-5.
short: F. Buchholz, L.M. Upterworth, L. Tueffers, E.E. Groth, K. Haas, D. Schütz,
A. Savietto Scholz, A. Batra, S. Pal, S. Banerjee, B.N. Dubey, S. Franzenburg,
B. Kalsdorf, K.F. Rabe, D. Nurjadi, J. Rupp, D.I. Andersson, H. Sondermann, M.
Bramkamp, R. Römhild, H. Schulenburg, Nature Communications 17 (2026).
corr_author: '1'
date_created: 2026-05-31T22:02:12Z
date_published: 2026-05-20T00:00:00Z
date_updated: 2026-06-02T07:14:35Z
day: '20'
ddc:
- '570'
department:
- _id: CaGu
doi: 10.1038/s41467-026-71178-5
file:
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checksum: aa29f8806908dc0469dff21e2d5ad01f
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creator: dernst
date_created: 2026-06-02T07:11:12Z
date_updated: 2026-06-02T07:11:12Z
file_id: '21936'
file_name: 2026_NatureComm_Buchholz.pdf
file_size: 1276166
relation: main_file
success: 1
file_date_updated: 2026-06-02T07:11:12Z
has_accepted_license: '1'
intvolume: ' 17'
language:
- iso: eng
month: '05'
oa: 1
oa_version: Published Version
project:
- _id: bd6f94d1-d553-11ed-ba76-ae9f07250f74
grant_number: E219
name: Non-canonical antibiotic interactions
publication: Nature Communications
publication_identifier:
eissn:
- 2041-1723
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Robust antibiotic sensitization of pathogenic Pseudomonas aeruginosa via negative
hysteresis in the cell envelope
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 17
year: '2026'
...
---
APC_amount: 5949 EUR
OA_place: publisher
OA_type: hybrid
_id: '19626'
abstract:
- lang: eng
text: Active regulation of gene expression, orchestrated by complex interactions
of activators and repressors at promoters, controls the fate of organisms. In
contrast, basal expression at uninduced promoters is considered to be a dynamically
inert mode of nonfunctional “promoter leakiness,” merely a byproduct of transcriptional
regulation. Here, we investigate the basal expression mode of the mar operon,
the main regulator of intrinsic multiple antibiotic resistance in Escherichia
coli, and link its dynamic properties to the noncanonical, yet highly conserved
start codon of marR across Enterobacteriaceae. Real-time, single-cell measurements
across tens of generations reveal that basal expression consists of rare stochastic
gene expression pulses, which maximize variability in wildtype and, surprisingly,
transiently accelerate cellular elongation rates. Competition experiments show
that basal expression confers fitness advantages to wildtype across several transitions
between exponential and stationary growth by shortening lag times. The dynamically
rich basal expression of the mar operon has likely been evolutionarily maintained
for its role in growth homeostasis of Enterobacteria within the gut environment,
thereby allowing other ancillary gene regulatory roles to evolve, e.g., control
of costly-to-induce multidrug efflux pumps. Understanding the complex selection
forces governing genetic systems involved in intrinsic multidrug resistance is
crucial for effective public health measures.
acknowledged_ssus:
- _id: Bio
acknowledgement: K.J. thanks B. Wu, I. Tomanek, K. Tomasek for detailed discussions
on the manuscript, all other members from the Guet laboratory for valuable feedback,
R. Chait, & Imaging and Optics Facility, Institute of Science and Technology Austria
for helping with microscopy, Dr. Sudha Rao and Dr. Raja Mugasimangalam, Genotypic
Technology India for allowing time off to address the revisions. K.J. acknowledges
Institute of Science and Technology fellowship IC1006FELL02, R.H. was supported
in part by Chan Zuckerberg Initiative and Donor Advised-Fund grant 2020-225401 (https://doi.org/10.37921/120055ratwvi),
O.O.B. acknowledges Fonds Zur Förderung der Wissenschaftlichen Forschung (FWF) Grant
ESP253-B, R.R. acknowledges FWF Grant 10.55776/ESP219, C.C.G. acknowledges FWF I5127-B.
article_number: e2413709122
article_processing_charge: Yes (in subscription journal)
article_type: original
author:
- first_name: Kirti
full_name: Jain, Kirti
id: 330F0278-F248-11E8-B48F-1D18A9856A87
last_name: Jain
orcid: 0000-0002-3809-0449
- first_name: Robert
full_name: Hauschild, Robert
id: 4E01D6B4-F248-11E8-B48F-1D18A9856A87
last_name: Hauschild
orcid: 0000-0001-9843-3522
- first_name: Olga
full_name: Bochkareva, Olga
id: C4558D3C-6102-11E9-A62E-F418E6697425
last_name: Bochkareva
orcid: 0000-0003-1006-6639
- first_name: Roderich
full_name: Römhild, Roderich
id: 68E56E44-62B0-11EA-B963-444F3DDC885E
last_name: Römhild
orcid: 0000-0001-9480-5261
- first_name: Gašper
full_name: Tkačik, Gašper
id: 3D494DCA-F248-11E8-B48F-1D18A9856A87
last_name: Tkačik
orcid: 0000-0002-6699-1455
- first_name: Calin C
full_name: Guet, Calin C
id: 47F8433E-F248-11E8-B48F-1D18A9856A87
last_name: Guet
orcid: 0000-0001-6220-2052
citation:
ama: Jain K, Hauschild R, Bochkareva O, Römhild R, Tkačik G, Guet CC. Pulsatile
basal gene expression as a fitness determinant in bacteria. Proceedings of
the National Academy of Sciences. 2025;122(15). doi:10.1073/pnas.2413709122
apa: Jain, K., Hauschild, R., Bochkareva, O., Römhild, R., Tkačik, G., & Guet,
C. C. (2025). Pulsatile basal gene expression as a fitness determinant in bacteria.
Proceedings of the National Academy of Sciences. National Academy of Sciences.
https://doi.org/10.1073/pnas.2413709122
chicago: Jain, Kirti, Robert Hauschild, Olga Bochkareva, Roderich Römhild, Gašper
Tkačik, and Calin C Guet. “Pulsatile Basal Gene Expression as a Fitness Determinant
in Bacteria.” Proceedings of the National Academy of Sciences. National
Academy of Sciences, 2025. https://doi.org/10.1073/pnas.2413709122.
ieee: K. Jain, R. Hauschild, O. Bochkareva, R. Römhild, G. Tkačik, and C. C. Guet,
“Pulsatile basal gene expression as a fitness determinant in bacteria,” Proceedings
of the National Academy of Sciences, vol. 122, no. 15. National Academy of
Sciences, 2025.
ista: Jain K, Hauschild R, Bochkareva O, Römhild R, Tkačik G, Guet CC. 2025. Pulsatile
basal gene expression as a fitness determinant in bacteria. Proceedings of the
National Academy of Sciences. 122(15), e2413709122.
mla: Jain, Kirti, et al. “Pulsatile Basal Gene Expression as a Fitness Determinant
in Bacteria.” Proceedings of the National Academy of Sciences, vol. 122,
no. 15, e2413709122, National Academy of Sciences, 2025, doi:10.1073/pnas.2413709122.
short: K. Jain, R. Hauschild, O. Bochkareva, R. Römhild, G. Tkačik, C.C. Guet, Proceedings
of the National Academy of Sciences 122 (2025).
corr_author: '1'
date_created: 2025-04-27T22:02:13Z
date_published: 2025-04-15T00:00:00Z
date_updated: 2026-05-20T08:33:08Z
day: '15'
ddc:
- '570'
department:
- _id: CaGu
- _id: Bio
- _id: FyKo
- _id: GaTk
doi: 10.1073/pnas.2413709122
external_id:
isi:
- '001471235200001'
pmid:
- '40193613'
file:
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checksum: 115a687f40009660eb4b38b4f6559d41
content_type: application/pdf
creator: dernst
date_created: 2025-06-24T07:27:43Z
date_updated: 2025-06-24T07:27:43Z
file_id: '19888'
file_name: 2025_PNAS_Jain.pdf
file_size: 2949523
relation: main_file
success: 1
file_date_updated: 2025-06-24T07:27:43Z
has_accepted_license: '1'
intvolume: ' 122'
isi: 1
issue: '15'
language:
- iso: eng
month: '04'
oa: 1
oa_version: Published Version
pmid: 1
project:
- _id: c08e9ad1-5a5b-11eb-8a69-9d1cf3b07473
grant_number: CZI01
name: Tools for automation and feedback microscopy
- _id: bd6f94d1-d553-11ed-ba76-ae9f07250f74
grant_number: E219
name: Non-canonical antibiotic interactions
- _id: 34e076d6-11ca-11ed-8bc3-aec76c41a181
grant_number: I05127
name: Evolutionary analysis of gene regulation
publication: Proceedings of the National Academy of Sciences
publication_identifier:
eissn:
- 1091-6490
issn:
- 0027-8424
publication_status: published
publisher: National Academy of Sciences
quality_controlled: '1'
related_material:
link:
- description: News on ISTA website
relation: press_release
url: https://ista.ac.at/en/news/clockwork-just-for-antibiotic-resistance/
record:
- id: '19294'
relation: research_data
status: public
scopus_import: '1'
status: public
title: Pulsatile basal gene expression as a fitness determinant in bacteria
tmp:
image: /images/cc_by.png
legal_code_url: https://creativecommons.org/licenses/by/4.0/legalcode
name: Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)
short: CC BY (4.0)
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 122
year: '2025'
...