Distinct requirements for tail-anchored membrane protein biogenesis in escherichia coli

Markus Peschke, Mélanie Le Goff, Gregory M. Koningstein, Norbert O. Vischer, Abbi Abdel-Rehim, Stephen High, Peter Van Ulsen, Joen Luirink

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Tail-anchored membrane proteins (TAMPs) are a distinct subset of inner membrane proteins (IMPs) characterized by a single C-terminal transmembrane domain (TMD) that is responsible for both targeting and anchoring. Little is known about the routing of TAMPs in bacteria. Here, we have investigated the role of TMD hydrophobicity in tail-anchor function in Escherichia coli and its influence on the choice of targeting/insertion pathway. We created a set of synthetic, fluorescent TAMPs that vary in the hydrophobicity of their TMDs and corresponding control polypeptides that are extended at their C terminus to create regular type II IMPs. Surprisingly, we observed that TAMPs have a much lower TMD hydrophobicity threshold for efficient targeting and membrane insertion than their type II counterparts. Using strains conditional for the expression of known membrane-targeting and insertion factors, we show that TAMPs with strongly hydrophobic TMDs require the signal recognition particle (SRP) for targeting. Neither the SecYEG translocon nor YidC appears to be essential for the membrane insertion of any of the TAMPs studied. In contrast, corresponding type II IMPs with a TMD of sufficient hydrophobicity to promote membrane insertion followed an SRP- and SecYEG translocon-dependent pathway. Together, these data indicate that the capacity of a TMD to promote the biogenesis of E. coli IMPs is strongly dependent upon the polypeptide context in which it is presented.

Original languageEnglish
Article numbere01580-19
JournalmBio
Volume10
Issue number5
DOIs
Publication statusPublished - 1 Jan 2019

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Membrane Proteins
Escherichia coli
Hydrophobic and Hydrophilic Interactions
Signal Recognition Particle
Membranes
Peptides
Bacteria

Keywords

  • Escherichia coli
  • Hydrophobicity
  • Membrane proteins
  • Membrane targeting
  • Tail-anchored

Cite this

Peschke, Markus ; Le Goff, Mélanie ; Koningstein, Gregory M. ; Vischer, Norbert O. ; Abdel-Rehim, Abbi ; High, Stephen ; Van Ulsen, Peter ; Luirink, Joen. / Distinct requirements for tail-anchored membrane protein biogenesis in escherichia coli. In: mBio. 2019 ; Vol. 10, No. 5.
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abstract = "Tail-anchored membrane proteins (TAMPs) are a distinct subset of inner membrane proteins (IMPs) characterized by a single C-terminal transmembrane domain (TMD) that is responsible for both targeting and anchoring. Little is known about the routing of TAMPs in bacteria. Here, we have investigated the role of TMD hydrophobicity in tail-anchor function in Escherichia coli and its influence on the choice of targeting/insertion pathway. We created a set of synthetic, fluorescent TAMPs that vary in the hydrophobicity of their TMDs and corresponding control polypeptides that are extended at their C terminus to create regular type II IMPs. Surprisingly, we observed that TAMPs have a much lower TMD hydrophobicity threshold for efficient targeting and membrane insertion than their type II counterparts. Using strains conditional for the expression of known membrane-targeting and insertion factors, we show that TAMPs with strongly hydrophobic TMDs require the signal recognition particle (SRP) for targeting. Neither the SecYEG translocon nor YidC appears to be essential for the membrane insertion of any of the TAMPs studied. In contrast, corresponding type II IMPs with a TMD of sufficient hydrophobicity to promote membrane insertion followed an SRP- and SecYEG translocon-dependent pathway. Together, these data indicate that the capacity of a TMD to promote the biogenesis of E. coli IMPs is strongly dependent upon the polypeptide context in which it is presented.",
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Distinct requirements for tail-anchored membrane protein biogenesis in escherichia coli. / Peschke, Markus; Le Goff, Mélanie; Koningstein, Gregory M.; Vischer, Norbert O.; Abdel-Rehim, Abbi; High, Stephen; Van Ulsen, Peter; Luirink, Joen.

In: mBio, Vol. 10, No. 5, e01580-19, 01.01.2019.

Research output: Contribution to JournalArticleAcademicpeer-review

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T1 - Distinct requirements for tail-anchored membrane protein biogenesis in escherichia coli

AU - Peschke, Markus

AU - Le Goff, Mélanie

AU - Koningstein, Gregory M.

AU - Vischer, Norbert O.

AU - Abdel-Rehim, Abbi

AU - High, Stephen

AU - Van Ulsen, Peter

AU - Luirink, Joen

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N2 - Tail-anchored membrane proteins (TAMPs) are a distinct subset of inner membrane proteins (IMPs) characterized by a single C-terminal transmembrane domain (TMD) that is responsible for both targeting and anchoring. Little is known about the routing of TAMPs in bacteria. Here, we have investigated the role of TMD hydrophobicity in tail-anchor function in Escherichia coli and its influence on the choice of targeting/insertion pathway. We created a set of synthetic, fluorescent TAMPs that vary in the hydrophobicity of their TMDs and corresponding control polypeptides that are extended at their C terminus to create regular type II IMPs. Surprisingly, we observed that TAMPs have a much lower TMD hydrophobicity threshold for efficient targeting and membrane insertion than their type II counterparts. Using strains conditional for the expression of known membrane-targeting and insertion factors, we show that TAMPs with strongly hydrophobic TMDs require the signal recognition particle (SRP) for targeting. Neither the SecYEG translocon nor YidC appears to be essential for the membrane insertion of any of the TAMPs studied. In contrast, corresponding type II IMPs with a TMD of sufficient hydrophobicity to promote membrane insertion followed an SRP- and SecYEG translocon-dependent pathway. Together, these data indicate that the capacity of a TMD to promote the biogenesis of E. coli IMPs is strongly dependent upon the polypeptide context in which it is presented.

AB - Tail-anchored membrane proteins (TAMPs) are a distinct subset of inner membrane proteins (IMPs) characterized by a single C-terminal transmembrane domain (TMD) that is responsible for both targeting and anchoring. Little is known about the routing of TAMPs in bacteria. Here, we have investigated the role of TMD hydrophobicity in tail-anchor function in Escherichia coli and its influence on the choice of targeting/insertion pathway. We created a set of synthetic, fluorescent TAMPs that vary in the hydrophobicity of their TMDs and corresponding control polypeptides that are extended at their C terminus to create regular type II IMPs. Surprisingly, we observed that TAMPs have a much lower TMD hydrophobicity threshold for efficient targeting and membrane insertion than their type II counterparts. Using strains conditional for the expression of known membrane-targeting and insertion factors, we show that TAMPs with strongly hydrophobic TMDs require the signal recognition particle (SRP) for targeting. Neither the SecYEG translocon nor YidC appears to be essential for the membrane insertion of any of the TAMPs studied. In contrast, corresponding type II IMPs with a TMD of sufficient hydrophobicity to promote membrane insertion followed an SRP- and SecYEG translocon-dependent pathway. Together, these data indicate that the capacity of a TMD to promote the biogenesis of E. coli IMPs is strongly dependent upon the polypeptide context in which it is presented.

KW - Escherichia coli

KW - Hydrophobicity

KW - Membrane proteins

KW - Membrane targeting

KW - Tail-anchored

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