Forces on Nascent Polypeptides during Membrane Insertion and Translocation via the Sec Translocon

Niesen, Michiel J.M.; Müller-Lucks, Annika; Hedman, Rickard; Heijne, Gunnar von; Miller, Thomas F.

doi:10.1016/j.bpj.2018.10.002

Cited by 32 publications

(44 citation statements)

References 40 publications

(86 reference statements)

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“…To evaluate the connection between pulling forces on the nascent chain and frameshifting, we measured the tension on the nascent chain at the point of elongation when the slip site occupies the ribosomal active site (Supplemental Movie 1). 41 Pulling forces were highest for the LL variant, which averaged 4.2 pN higher than the WT. In contrast, the EE mutations reduce the pulling force on the nascent chain by an average 2.3 pN relative to WT.…”

Section: Impact Of Nascent Chain Forces On Ribosomal Frameshiftingmentioning

confidence: 92%

“…This finding suggests pulling forces are generated by the movement of hydrophobic transmembrane segments from the protein-conducting channel of the translocon to the hydrophobic membrane core, as has been established previously. 34,35,41 This interpretation suggests differences in pulling forces arise from variations in the distribution of topological isomers that form during translation of these variants (Fig. 5D).…”

Section: Impact Of Nascent Chain Forces On Ribosomal Frameshiftingmentioning

confidence: 94%

“…39 These simulations were previously found to sufficiently recapitulate several aspects of cotranslational membrane protein folding including the formation of topological isomers and the generation of tension on the nascent chain. 38,40,41 CGMD simulations of SINV polyprotein biosynthesis suggest the nascent chain samples several different topological isomers (Fig. 5A), and that its topological heterogeneity persists after the polyprotein has cleared the translocon.…”

Section: Impact Of Nascent Chain Forces On Ribosomal Frameshiftingmentioning

confidence: 99%

“…Despite the significant simplifications involved in this model, the CGMD model has proven accurate in capturing the integration probabilities, topology distributions, and forces experienced in previous studies. 38,39,41 In order to obtain the distribution of topologies for various polyprotein mutants, the translation and integration of each sequence was simulated 560 times. In order to reduce computational cost, simulations only included the first three TMDs of the alphavirus polyprotein.…”

Section: Course Grained Simulations Of Polyprotein Translationmentioning

confidence: 99%

“…This protocol is analogous to force measurements performed in previous work, with exception of a shortened pausing duration. 41 The datasets described herein are available from the corresponding authors upon request.…”

Section: Course Grained Simulations Of Polyprotein Translationmentioning

confidence: 99%

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Cotranslational Folding Stimulates Programmed Ribosomal Frameshifting in the Alphavirus Structural Polyprotein

Harrington

Zimmer

Chamness

et al. 2019

Preprint

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Viruses maximize their genetic coding capacity through a variety of biochemical mechanisms including programmed ribosomal frameshifting (PRF), which facilitates the production of multiple proteins from a single transcript. PRF is typically stimulated by structural elements within the mRNA that generate mechanical tension between the transcript and ribosome. However, in this work we show that the forces generated by the cotranslational folding of the nascent polypeptide chain can also enhance PRF. Using an array of biochemical, cellular, and computational techniques, we first demonstrate that the Sindbis virus structural polyprotein forms two competing topological isomers during biosynthesis at the ribosome-translocon complex. We then show that the formation of one of these topological isomers is linked to PRF. Coarse-grained molecular dynamic simulations reveal that the translocon-mediated membrane integration of a transmembrane domain upstream from the ribosomal slip-site generates a force on the nascent polypeptide chain that scales with observed frameshifting. Together, our results demonstrate that cotranslational folding of this protein generates a tension that stimulates PRF. To our knowledge, this constitutes the first example in which the conformational state of the nascent chain has been linked to PRF. These findings raise the possibility that, in addition to RNA-mediated translational recoding, a variety of cotranslational folding and/ or binding events may also stimulate PRF.

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Section: Impact Of Nascent Chain Forces On Ribosomal Frameshiftingmentioning

confidence: 92%

Section: Impact Of Nascent Chain Forces On Ribosomal Frameshiftingmentioning

confidence: 94%

Section: Impact Of Nascent Chain Forces On Ribosomal Frameshiftingmentioning

confidence: 99%

Section: Course Grained Simulations Of Polyprotein Translationmentioning

confidence: 99%

Section: Course Grained Simulations Of Polyprotein Translationmentioning

confidence: 99%

See 3 more Smart Citations

Cotranslational Folding Stimulates Programmed Ribosomal Frameshifting in the Alphavirus Structural Polyprotein

Harrington

Zimmer

Chamness

et al. 2019

Preprint

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Upstream charged and hydrophobic residues impact the timing of membrane insertion of transmembrane helices

et al. 2022

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During SecYEG‐mediated cotranslational insertion of membrane proteins, transmembrane helices (TMHs) first make contact with the membrane when their N‐terminal end is ~ 45 residues away from the peptidyl transferase centre. However, we recently uncovered instances where the first contact is delayed by up to ~ 10 residues. Here, we recapitulate these effects using a model TMH fused to two short segments from the Escherichia coli inner membrane protein BtuC: a positively charged loop and a re‐entrant loop. We show that the critical residues are two Arg residues in the positively charged loop and four hydrophobic residues in the re‐entrant loop. Thus, both electrostatic and hydrophobic interactions involving sequence elements that are not part of a TMH can impact the way the latter behaves during membrane insertion.

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Structural insight into co-translational membrane protein folding

Pellowe

Booth

2020

Biochimica et Biophysica Acta (BBA) - Biomembranes

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Forces on Nascent Polypeptides during Membrane Insertion and Translocation via the Sec Translocon

Cited by 32 publications

References 40 publications

Cotranslational Folding Stimulates Programmed Ribosomal Frameshifting in the Alphavirus Structural Polyprotein

Cotranslational Folding Stimulates Programmed Ribosomal Frameshifting in the Alphavirus Structural Polyprotein

Upstream charged and hydrophobic residues impact the timing of membrane insertion of transmembrane helices

Structural insight into co-translational membrane protein folding

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