# Investigation of multifunctional proteins that integrate packaging RNPs, RNA export, and translation

> **NIH NIH R15** · WAKE FOREST UNIVERSITY · 2020 · $430,122

## Abstract

Project Summary
Eukaryotic gene expression depends on many steps in both the nucleus and cytoplasm. In the
nucleus, large nascent RNAs, such as ribosomal RNAs and mRNAs, are manufactured and
assembled into ribonucleoprotein particles (RNPs) that are exported to the cytoplasm for protein
synthesis. The proteins involved in RNP assembly and export play critical roles throughout gene
expression from co-transcriptional RNA processing to translation.
 AAA+ ATPases are a large and functionally diverse family of proteins that use the energy of
ATP binding and hydrolysis to induce conformational changes and remodeling in various protein
substrates. Loss of Elf1 (Elongation-Like Factor 1), an AAA+ superfamily ATPase implicated in
RNA nuclear export, causes severe growth defects that can be mitigated by spontaneous
suppressor mutations.
 We confirmed and isolated two suppressor mutants: an endonuclease, Cue2, and a large
ribosomal subunit protein, Rpl2702. Elf1 co-purifies with these mutants, providing additional
support for their functional connection. Using affinity purification and mass spectrometry
analysis of Elf1 and Cue2, we have developed a molecular framework to systematically
investigate their roles in the multifaceted regulation of posttranscriptional gene expression from
RNP export to translation to ribosome-associated quality control. In addition, we have observed
RNA export defects with loss of Elf1. The resulting nuclear retention of RNA destabilizes the
genome, probably because abnormal DNA-RNA hybrids (R-loops) form.
 We hypothesize that Elf1 is associated with RNPs and functions in RNA/ribosome export
and translation in different cellular compartments, antagonistically regulated by Cue2. To test
this hypothesis, we will investigate the integrated roles of Elf1, Cue2, and Rpl2702 in
maintaining genome stability (Aim 1), RNA and/or ribosome nuclear export (Aim 2), and
translation elongation and ribosome-associated quality control (Aim 3). We will use traditional
molecular biology and biochemical approaches, and also develop new genetic tools to analyze
transcription-dependent hyper-recombination and examine various types of ribosome-
associated mRNA decays. Our hypotheses and research strategy are based on a host of
preliminary findings. Results are expected to advance the fields of RNA biology, protein
synthesis, and genomic instability. Through implementation of research and student training, we
will generate new quantitative genetic tools that will be appreciated in the fission yeast
community.

## Key facts

- **NIH application ID:** 10047135
- **Project number:** 1R15GM139107-01
- **Recipient organization:** WAKE FOREST UNIVERSITY
- **Principal Investigator:** Ke Zhang Reid
- **Activity code:** R15 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $430,122
- **Award type:** 1
- **Project period:** 2020-09-01 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10047135

## Citation

> US National Institutes of Health, RePORTER application 10047135, Investigation of multifunctional proteins that integrate packaging RNPs, RNA export, and translation (1R15GM139107-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10047135. Licensed CC0.

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