The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity

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The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity. / Jamshad, Mohammed; Knowles, Timothy J; White, Scott A; Ward, Douglas G; Mohammed, Fiyaz; Rahman, Kazi Fahmida; Wynne, Max; Hughes, Gareth W; Kramer, Günter; Bukau, Bernd; Huber, Damon.

In: Elife, Vol. 8, e48385, 27.06.2019.

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@article{05bf9bf0bb46468693eb1b11d0e28a00,
title = "The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity",
abstract = "In bacteria, the translocation of proteins across the cytoplasmic membrane by the Sec machinery requires the ATPase SecA. SecA binds ribosomes and recognises nascent substrate proteins, but the molecular mechanism of nascent substrate recognition is unknown. We investigated the role of the C-terminal tail (CTT) of SecA in nascent polypeptide recognition. The CTT consists of a flexible linker (FLD) and a small metal-binding domain (MBD). Phylogenetic analysis and ribosome binding experiments indicated that the MBD interacts with 70S ribosomes. Disruption of the MBD only or the entire CTT had opposing effects on ribosome binding, substrate-protein binding, ATPase activity and in vivo function, suggesting that the CTT influences the conformation of SecA. Site-specific crosslinking indicated that F399 in SecA contacts ribosomal protein uL29, and binding to nascent chains disrupts this interaction. Structural studies provided insight into the CTT-mediated conformational changes in SecA. Our results suggest a mechanism for nascent substrate protein recognition.",
keywords = "E. coli, SAXS, SecA, biochemistry, chemical biology, metal-binding domain, molecular biophysics, protein secretion, protein translocation, ribosome, structural biology",
author = "Mohammed Jamshad and Knowles, {Timothy J} and White, {Scott A} and Ward, {Douglas G} and Fiyaz Mohammed and Rahman, {Kazi Fahmida} and Max Wynne and Hughes, {Gareth W} and G{\"u}nter Kramer and Bernd Bukau and Damon Huber",
year = "2019",
month = "6",
day = "27",
doi = "10.7554/eLife.48385.001",
language = "English",
volume = "8",
journal = "Elife",
issn = "2050-084X",
publisher = "eLife Sciences Publications",

}

RIS

TY - JOUR

T1 - The C-terminal tail of the bacterial translocation ATPase SecA modulates its activity

AU - Jamshad, Mohammed

AU - Knowles, Timothy J

AU - White, Scott A

AU - Ward, Douglas G

AU - Mohammed, Fiyaz

AU - Rahman, Kazi Fahmida

AU - Wynne, Max

AU - Hughes, Gareth W

AU - Kramer, Günter

AU - Bukau, Bernd

AU - Huber, Damon

PY - 2019/6/27

Y1 - 2019/6/27

N2 - In bacteria, the translocation of proteins across the cytoplasmic membrane by the Sec machinery requires the ATPase SecA. SecA binds ribosomes and recognises nascent substrate proteins, but the molecular mechanism of nascent substrate recognition is unknown. We investigated the role of the C-terminal tail (CTT) of SecA in nascent polypeptide recognition. The CTT consists of a flexible linker (FLD) and a small metal-binding domain (MBD). Phylogenetic analysis and ribosome binding experiments indicated that the MBD interacts with 70S ribosomes. Disruption of the MBD only or the entire CTT had opposing effects on ribosome binding, substrate-protein binding, ATPase activity and in vivo function, suggesting that the CTT influences the conformation of SecA. Site-specific crosslinking indicated that F399 in SecA contacts ribosomal protein uL29, and binding to nascent chains disrupts this interaction. Structural studies provided insight into the CTT-mediated conformational changes in SecA. Our results suggest a mechanism for nascent substrate protein recognition.

AB - In bacteria, the translocation of proteins across the cytoplasmic membrane by the Sec machinery requires the ATPase SecA. SecA binds ribosomes and recognises nascent substrate proteins, but the molecular mechanism of nascent substrate recognition is unknown. We investigated the role of the C-terminal tail (CTT) of SecA in nascent polypeptide recognition. The CTT consists of a flexible linker (FLD) and a small metal-binding domain (MBD). Phylogenetic analysis and ribosome binding experiments indicated that the MBD interacts with 70S ribosomes. Disruption of the MBD only or the entire CTT had opposing effects on ribosome binding, substrate-protein binding, ATPase activity and in vivo function, suggesting that the CTT influences the conformation of SecA. Site-specific crosslinking indicated that F399 in SecA contacts ribosomal protein uL29, and binding to nascent chains disrupts this interaction. Structural studies provided insight into the CTT-mediated conformational changes in SecA. Our results suggest a mechanism for nascent substrate protein recognition.

KW - E. coli

KW - SAXS

KW - SecA

KW - biochemistry

KW - chemical biology

KW - metal-binding domain

KW - molecular biophysics

KW - protein secretion

KW - protein translocation

KW - ribosome

KW - structural biology

U2 - 10.7554/eLife.48385.001

DO - 10.7554/eLife.48385.001

M3 - Article

C2 - 31246174

VL - 8

JO - Elife

JF - Elife

SN - 2050-084X

M1 - e48385

ER -