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Cotranslational Protein Translocation

Cotranslational Protein Translocation

Translocation of proteins across membranes is an ancient process that occurs even in bacteria and archaebacteria. In fact, the components of the translocation machinery are still conserved between prokaryotes and eukaryotes.
Sec61 channel partners for cotranslational translocation
During cotranslational translocation, the Sec61 channel partners with the signal recognition particle (SRP), the signal recognition particle receptor (SR), and the ribosomes to transport the nascent polypeptide chain...

Regulated mRNA Transport

Regulated mRNA Transport

In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...

Regulated mRNA Transport

Regulated mRNA Transport

In eukaryotes, transcription and translation are compartmentalized; an mRNA is first synthesized in the nucleus and then selectively transported to the cytoplasm for protein synthesis. Before transport, a pre-mRNA undergoes several steps of post-transcriptional modifications including splicing, 5' capping, and the addition of a poly-adenine tail. Various proteins bind to the pre-mRNA during these modifications. The mRNA transport takes place with the help of multiple proteins playing specific...

Post-translational Translocation of Proteins to the RER

Post-translational Translocation of Proteins to the RER

A sizable fraction of proteins destined for ER are first synthesized in the cell cytosol and then transported across the ER membrane–a process called post-translational translocation. Similar to cotranslationally translocated proteins, these proteins also use the Sec translocon complex to enter the ER lumen.
Targeting proteins to the ER
Hsp40 and Hsp70 chaperone molecules bind the translated proteins in the cytosol to prevent their folding. The chaperone binding helps to keep the signal...

Initiation of Translation

Initiation of Translation

Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...

Initiation of Translation

Initiation of Translation

Initiating translation is complex because it involves multiple molecules. Initiator tRNA, ribosomal subunits, and eukaryotic initiation factors (eIFs) are all required to assemble on the initiation codon of mRNA. This process consists of several steps that are mediated by different eIFs.
First, the initiator tRNA must be selected from the pool of elongator tRNAs by eukaryotic initiation factor 2 (eIF2). The initiator tRNA (Met-tRNAi) has conserved sequence elements including modified bases at...

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Related Experiment Video

Updated: May 19, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

Published on: February 25, 2011

Intermediate states during mRNA-tRNA translocation.

Joachim Frank¹

¹Howard Hughes Medical Institute, Department of Biochemistry and Molecular Biophysics, Columbia University, 650 W. 168th Street, New York, NY 10032, United States. jf2192@columbia.edu

Current Opinion in Structural Biology

|August 22, 2012

Summary

The ribosomal complex acts as a Brownian machine, exploring various states. Elongation Factor G (EF-G) binding stabilizes a key state, driving the translocation process.

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Last Updated: May 19, 2026

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses

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Published on: February 25, 2011

Dual DNA Rulers to Study the Mechanism of Ribosome Translocation with Single-Nucleotide Resolution

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Published on: July 8, 2019

Isolation of mRNAs Associated with Yeast Mitochondria to Study Mechanisms of Localized Translation

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Published on: March 14, 2014

Area of Science:

Molecular Biology
Biophysics
Structural Biology

Background:

The pre-translocation (PRE) ribosomal complex exhibits dynamic behavior.
This complex fluctuates between multiple conformations and transfer RNA (tRNA) binding states.

Purpose of the Study:

To investigate the conformational dynamics of the PRE ribosomal complex.
To elucidate the role of intermediate states in ribosomal translocation.

Main Methods:

Cryogenic-electron microscopy (cryo-EM)
X-ray crystallography
Single-molecule fluorescence resonance energy transfer (smFRET)

Main Results:

Identified multiple intermediate conformational states of the PRE complex.
Demonstrated that the PRE complex functions as a Brownian machine.
Showed that Elongation Factor G (EF-G) binding stabilizes the productive macrostate II (MS II).

Conclusions:

Ribosomal translocation initiates as a Brownian process.
The binding of EF-G is crucial for stabilizing the MS II conformation, facilitating translocation.