Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases

Research output: Contribution to journalArticlepeer-review

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Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases. / Lang, Silvia; Gruber, Karl; Mihajlovic, Sanja; Arnold, Roland; Gruber, Christian J; Steinlechner, Sonja; Jehl, Marc-Andre; Rattei, Thomas; Fröhlich, Kai-Uwe; Zechner, Ellen L.

In: Molecular Microbiology, Vol. 78, No. 6, 12.2010, p. 1539-55.

Research output: Contribution to journalArticlepeer-review

Harvard

Lang, S, Gruber, K, Mihajlovic, S, Arnold, R, Gruber, CJ, Steinlechner, S, Jehl, M-A, Rattei, T, Fröhlich, K-U & Zechner, EL 2010, 'Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases', Molecular Microbiology, vol. 78, no. 6, pp. 1539-55. https://doi.org/10.1111/j.1365-2958.2010.07423.x

APA

Lang, S., Gruber, K., Mihajlovic, S., Arnold, R., Gruber, C. J., Steinlechner, S., Jehl, M-A., Rattei, T., Fröhlich, K-U., & Zechner, E. L. (2010). Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases. Molecular Microbiology, 78(6), 1539-55. https://doi.org/10.1111/j.1365-2958.2010.07423.x

Vancouver

Author

Lang, Silvia ; Gruber, Karl ; Mihajlovic, Sanja ; Arnold, Roland ; Gruber, Christian J ; Steinlechner, Sonja ; Jehl, Marc-Andre ; Rattei, Thomas ; Fröhlich, Kai-Uwe ; Zechner, Ellen L. / Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases. In: Molecular Microbiology. 2010 ; Vol. 78, No. 6. pp. 1539-55.

Bibtex

@article{98539e6ea2e84b8a8d59651ee78abe2f,
title = "Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases",
abstract = "In preparation for transfer conjugative type IV secretion systems (T4SS) produce a nucleoprotein adduct containing a relaxase enzyme covalently linked to the 5' end of single-stranded plasmid DNA. The bound relaxase is expected to present features necessary for selective recognition by the type IV coupling protein (T4CP), which controls substrate entry to the envelope spanning secretion machinery. We prove that the IncF plasmid R1 relaxase TraI is translocated to the recipient cells. Using a Cre recombinase assay (CRAfT) we mapped two internally positioned translocation signals (TS) on F-like TraI proteins that independently mediate efficient recognition and secretion. Tertiary structure predictions for the TS matched best helicase RecD2 from Deinococcus radiodurans. The TS is widely conserved in MOB(F) and MOB(Q) families of relaxases. Structure/function relationships within the TS were identified by mutation. A key residue in specific recognition by T4CP TraD was revealed by a fidelity switch phenotype for an F to plasmid R1 exchange L626H mutation. Finally, we show that physical linkage of the relaxase catalytic domain to a TraI TS is necessary for efficient conjugative transfer.",
keywords = "Conjugation, Genetic, DNA Helicases, Escherichia coli, Escherichia coli Proteins, Multigene Family, Protein Sorting Signals, Protein Structure, Tertiary, Protein Transport, Journal Article, Research Support, Non-U.S. Gov't",
author = "Silvia Lang and Karl Gruber and Sanja Mihajlovic and Roland Arnold and Gruber, {Christian J} and Sonja Steinlechner and Marc-Andre Jehl and Thomas Rattei and Kai-Uwe Fr{\"o}hlich and Zechner, {Ellen L}",
note = "{\textcopyright} 2010 Blackwell Publishing Ltd.",
year = "2010",
month = dec,
doi = "10.1111/j.1365-2958.2010.07423.x",
language = "English",
volume = "78",
pages = "1539--55",
journal = "Molecular Microbiology",
issn = "0950-382X",
publisher = "Wiley",
number = "6",

}

RIS

TY - JOUR

T1 - Molecular recognition determinants for type IV secretion of diverse families of conjugative relaxases

AU - Lang, Silvia

AU - Gruber, Karl

AU - Mihajlovic, Sanja

AU - Arnold, Roland

AU - Gruber, Christian J

AU - Steinlechner, Sonja

AU - Jehl, Marc-Andre

AU - Rattei, Thomas

AU - Fröhlich, Kai-Uwe

AU - Zechner, Ellen L

N1 - © 2010 Blackwell Publishing Ltd.

PY - 2010/12

Y1 - 2010/12

N2 - In preparation for transfer conjugative type IV secretion systems (T4SS) produce a nucleoprotein adduct containing a relaxase enzyme covalently linked to the 5' end of single-stranded plasmid DNA. The bound relaxase is expected to present features necessary for selective recognition by the type IV coupling protein (T4CP), which controls substrate entry to the envelope spanning secretion machinery. We prove that the IncF plasmid R1 relaxase TraI is translocated to the recipient cells. Using a Cre recombinase assay (CRAfT) we mapped two internally positioned translocation signals (TS) on F-like TraI proteins that independently mediate efficient recognition and secretion. Tertiary structure predictions for the TS matched best helicase RecD2 from Deinococcus radiodurans. The TS is widely conserved in MOB(F) and MOB(Q) families of relaxases. Structure/function relationships within the TS were identified by mutation. A key residue in specific recognition by T4CP TraD was revealed by a fidelity switch phenotype for an F to plasmid R1 exchange L626H mutation. Finally, we show that physical linkage of the relaxase catalytic domain to a TraI TS is necessary for efficient conjugative transfer.

AB - In preparation for transfer conjugative type IV secretion systems (T4SS) produce a nucleoprotein adduct containing a relaxase enzyme covalently linked to the 5' end of single-stranded plasmid DNA. The bound relaxase is expected to present features necessary for selective recognition by the type IV coupling protein (T4CP), which controls substrate entry to the envelope spanning secretion machinery. We prove that the IncF plasmid R1 relaxase TraI is translocated to the recipient cells. Using a Cre recombinase assay (CRAfT) we mapped two internally positioned translocation signals (TS) on F-like TraI proteins that independently mediate efficient recognition and secretion. Tertiary structure predictions for the TS matched best helicase RecD2 from Deinococcus radiodurans. The TS is widely conserved in MOB(F) and MOB(Q) families of relaxases. Structure/function relationships within the TS were identified by mutation. A key residue in specific recognition by T4CP TraD was revealed by a fidelity switch phenotype for an F to plasmid R1 exchange L626H mutation. Finally, we show that physical linkage of the relaxase catalytic domain to a TraI TS is necessary for efficient conjugative transfer.

KW - Conjugation, Genetic

KW - DNA Helicases

KW - Escherichia coli

KW - Escherichia coli Proteins

KW - Multigene Family

KW - Protein Sorting Signals

KW - Protein Structure, Tertiary

KW - Protein Transport

KW - Journal Article

KW - Research Support, Non-U.S. Gov't

U2 - 10.1111/j.1365-2958.2010.07423.x

DO - 10.1111/j.1365-2958.2010.07423.x

M3 - Article

C2 - 21143323

VL - 78

SP - 1539

EP - 1555

JO - Molecular Microbiology

JF - Molecular Microbiology

SN - 0950-382X

IS - 6

ER -