# Time-resolved Cryo-EM of Short-lived States in Eukaryotic Translation

> **NIH NIH R01** · COLUMBIA UNIVERSITY HEALTH SCIENCES · 2020 · $346,241

## Abstract

Project Summary/Abstract
The overall objective of this research project is the elucidation of the process of translation (i.e., translation from
genetic information into protein) in eukaryotes, including mammals. Although X-ray structures exist for a number
of eukaryotic ribosomes, the structural basis of this process has been mainly investigated by cryo-electron
microscopy (cryo-EM). However, many short-lived states, with a life time of less than a second, cannot be
imaged by standard cryo-EM (characterized by pipetting/blotting of sample), leaving large gaps in our
understanding of these basic processes of life and in the knowledge base important for molecular medicine. In
the approach of time-resolved cryo-EM adopted in this lab, reactions are started in a microfluidic chip (silicon- or
plastic-based) by mixing two components, letting them react for a defined time (10 to 1000 ms) determined by
flow rate and length of reaction channel, and the reaction product is sprayed on the EM grid immediately before
the latter is plunged into the cryogen (liquid ethane, on the temperature of liquid nitrogen). In the current renewal
period of this grant, ending on March 31, 2019, this technique has been greatly improved, and applied to three
processes important in bacterial translation: initiation, termination, and ribosome recycling. All three applications
have been successful, resulting in the capture of a short-lived state at close-to-atomic resolution and leading to
papers either published or under consideration. For the current renewal, exploration of short-lived states in
eukaryotic translation will have even greater relevance for human health. In collaborations with leading experts
in eukaryotic translation, the Frank Lab team will apply time-resolved cryo-EM to the elucidation of processes
during eukaryotic translation initiation, termination, and recycling. The aim to determine atomic structures for
short-lived states that are impossible to capture by standard cryo-EM. Using these structures along with those
obtained from conventional cryo-EM and X-ray crystallography, the time courses and pathways taken in the
respective processes can be modeled for the first time, based on solid experiments. This new knowledge will
advance strategies for combatting many diseases that implicate dysfunctions of eukaryotic translation.

## Key facts

- **NIH application ID:** 10016331
- **Project number:** 5R01GM055440-23
- **Recipient organization:** COLUMBIA UNIVERSITY HEALTH SCIENCES
- **Principal Investigator:** JOACHIM FRANK
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $346,241
- **Award type:** 5
- **Project period:** 1997-01-01 → 2021-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10016331

## Citation

> US National Institutes of Health, RePORTER application 10016331, Time-resolved Cryo-EM of Short-lived States in Eukaryotic Translation (5R01GM055440-23). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10016331. Licensed CC0.

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