Different roles of conserved tyrosine residues of the acylated
domains in folding and activity of RTX toxins

J 2021

Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins

LEPESHEVA, A., A. OSICKOVA, J. HOLUBOVA, D. JURNECKA, S. KNOBLOCHOVA et. al.

Basic information

Original name

Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins

Authors

LEPESHEVA, A., A. OSICKOVA, J. HOLUBOVA, D. JURNECKA, S. KNOBLOCHOVA, C. ESPINOSA-VINALS, L. BUMBA, K. SKOPOVA, R. FISER, R. OSICKA, P. SEBO and J. MASIN

Edition

SCIENTIFIC REPORTS, 2021, 2045-2322

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10608 Biochemistry and molecular biology

Country of publisher

Germany

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 4.996

RIV identification code

RIV/00216224:14740/21:00124531

Organization unit

Central European Institute of Technology

DOI

http://dx.doi.org/10.1038/s41598-021-99112-3

UT WoS

000706380800096

Keywords in English

ADENYLATE-CYCLASE TOXINESCHERICHIA-COLI HEMOLYSINBORDETELLA-PERTUSSIS CYAACELL-INVASIVE ACTIVITYALPHA-HEMOLYSINMEMBRANE TRANSLOCATIONCOMPLEMENT RECEPTOR-3FATTY-ACYLATIONCALCIUMBINDING

Abstract

V originále

Pore-forming repeats in toxins (RTX) are key virulence factors of many Gram-negative pathogens. We have recently shown that the aromatic side chain of the conserved tyrosine residue 940 within the acylated segment of the RTX adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) plays a key role in target cell membrane interaction of the toxin. Therefore, we used a truncated CyaA-derived RTX719 construct to analyze the impact of Y940 substitutions on functional folding of the acylated segment of CyaA. Size exclusion chromatography combined with CD spectroscopy revealed that replacement of the aromatic side chain of Y940 by the side chains of alanine or proline residues disrupted the calcium-dependent folding of RTX719 and led to self-aggregation of the otherwise soluble and monomeric protein. Intriguingly, corresponding alanine substitutions of the conserved Y642, Y643 and Y639 residues in the homologous RtxA, HlyA and ApxIA hemolysins from Kingella kingae, Escherichia coli and Actinobacillus pleuropneumoniae, affected the membrane insertion, pore-forming (hemolytic) and cytotoxic capacities of these toxins only marginally. Activities of these toxins were impaired only upon replacement of the conserved tyrosines by proline residues. It appears, hence, that the critical role of the aromatic side chain of the Y940 residue is highly specific for the functional folding of the acylated domain of CyaA and determines its capacity to penetrate target cell membrane.

Links

90127, large research infrastructures

Name: CIISB II

Citovat

LEPESHEVA, A., A. OSICKOVA, J. HOLUBOVA, D. JURNECKA, S. KNOBLOCHOVA, C. ESPINOSA-VINALS, L. BUMBA, K. SKOPOVA, R. FISER, R. OSICKA, P. SEBO and J. MASIN. Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins. SCIENTIFIC REPORTS. 2021, vol. 11, No 1, p. 19814-19829. ISSN 2045-2322. Available from: https://dx.doi.org/10.1038/s41598-021-99112-3.

@article{1846841,
   author = {Lepesheva, A. and Osickova, A. and Holubova, J. and Jurnecka, D. and Knoblochova, S. and EspinosaandVinals, C. and Bumba, L. and Skopova, K. and Fiser, R. and Osicka, R. and Sebo, P. and Masin, J.},
   article_number = {1},
   doi = {http://dx.doi.org/10.1038/s41598-021-99112-3},
   keywords = {ADENYLATE-CYCLASE TOXINESCHERICHIA-COLI HEMOLYSINBORDETELLA-PERTUSSIS CYAACELL-INVASIVE ACTIVITYALPHA-HEMOLYSINMEMBRANE TRANSLOCATIONCOMPLEMENT RECEPTOR-3FATTY-ACYLATIONCALCIUMBINDING},
   language = {eng},
   issn = {2045-2322},
   journal = {SCIENTIFIC REPORTS},
   title = {Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins},
   url = {https://www.nature.com/articles/s41598-021-99112-3.pdf},
   volume = {11},
   year = {2021}
}

TY  - JOUR
ID  - 1846841
AU  - Lepesheva, A. - Osickova, A. - Holubova, J. - Jurnecka, D. - Knoblochova, S. - Espinosa-Vinals, C. - Bumba, L. - Skopova, K. - Fiser, R. - Osicka, R. - Sebo, P. - Masin, J.
PY  - 2021
TI  - Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins
JF  - SCIENTIFIC REPORTS
VL  - 11
IS  - 1
SP  - 19814
EP  - 19814
SN  - 20452322
KW  - ADENYLATE-CYCLASE TOXINESCHERICHIA-COLI HEMOLYSINBORDETELLA-PERTUSSIS CYAACELL-INVASIVE ACTIVITYALPHA-HEMOLYSINMEMBRANE TRANSLOCATIONCOMPLEMENT RECEPTOR-3FATTY-ACYLATIONCALCIUMBINDING
UR  - https://www.nature.com/articles/s41598-021-99112-3.pdf
N2  - Pore-forming repeats in toxins (RTX) are key virulence factors of many Gram-negative pathogens. We have recently shown that the aromatic side chain of the conserved tyrosine residue 940 within the acylated segment of the RTX adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) plays a key role in target cell membrane interaction of the toxin. Therefore, we used a truncated CyaA-derived RTX719 construct to analyze the impact of Y940 substitutions on functional folding of the acylated segment of CyaA. Size exclusion chromatography combined with CD spectroscopy revealed that replacement of the aromatic side chain of Y940 by the side chains of alanine or proline residues disrupted the calcium-dependent folding of RTX719 and led to self-aggregation of the otherwise soluble and monomeric protein. Intriguingly, corresponding alanine substitutions of the conserved Y642, Y643 and Y639 residues in the homologous RtxA, HlyA and ApxIA hemolysins from Kingella kingae, Escherichia coli and Actinobacillus pleuropneumoniae, affected the membrane insertion, pore-forming (hemolytic) and cytotoxic capacities of these toxins only marginally. Activities of these toxins were impaired only upon replacement of the conserved tyrosines by proline residues. It appears, hence, that the critical role of the aromatic side chain of the Y940 residue is highly specific for the functional folding of the acylated domain of CyaA and determines its capacity to penetrate target cell membrane.
ER  -

LEPESHEVA, A., A. OSICKOVA, J. HOLUBOVA, D. JURNECKA, S. KNOBLOCHOVA, C. ESPINOSA-VINALS, L. BUMBA, K. SKOPOVA, R. FISER, R. OSICKA, P. SEBO and J. MASIN. Different roles of conserved tyrosine residues of the acylated domains in folding and activity of RTX toxins. \textit{SCIENTIFIC REPORTS}. 2021, vol.~11, No~1, p.~19814-19829. ISSN~2045-2322. Available from: https://dx.doi.org/10.1038/s41598-021-99112-3.

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