# Molecular analysis of nuclear lamin assembly

> **NIH NIH F32** · CARNEGIE INSTITUTION OF WASHINGTON, D.C. · 2022 · $67,582

## Abstract

Project Summary
Lamin filaments are central structural organizers of the metazoan nucleus. They contribute to nuclear function
by controlling nuclear structure, separating the nucleoplasm from the cytoplasm and organizing the genome into
differentially regulated subdomains. Many diseases are associated with lamin dysregulation and abnormal
nuclear structure, underscoring the importance of these molecules. Despite the importance of lamins in normal
nuclear function, molecular mechanisms controlling lamin assembly are poorly understood. Previous studies
attempted to dissect lamin assembly using recombinant lamin proteins purified under denaturing conditions and
simultaneously refolded and assembled into filamentous structures through removal of denaturant. It is now clear
that the lamin structures assembled in these experiments do not resemble lamin filaments in cells. The goal of
this proposal is to develop experimental systems for studying physiological lamin assembly and to determine the
assembly pathway and mechanism of lamin assembly. The research is expected to extend understanding of
nuclear structure and function, and of diseases associated with dysregulation of nuclear structure and function.
 The proposed experiments aim to uncover molecular mechanisms of lamin assembly. In vitro
experiments will be conducted in Xenopus laevis egg extracts, which contain their own soluble lamin protein,
eliminating the need for recombinant lamins in assembly assays. Xenopus egg extracts can assemble diverse
lamin structures. By varying assembly conditions and studying these lamin assemblies using fluorescence and
electron microscopy, cellular structures and signals that control lamin assembly will be identified. Using analytical
biochemistry, the soluble lamin subunit will be characterized, along with any proteins that are in a stable complex
with the soluble lamin subunit. Importin  and  are known binding partners of soluble lamin in Xenopus egg
extract. Proposed experiments will determine how importins and other lamin-binding proteins regulate lamin
assembly. In vivo experiments will be conducted in genome edited stem cells. Mouse embryonic stem cells with
the genes encoding all three lamin isoforms knocked out have been isolated and propagated by the host lab.
Inducibly expressing fluorescently tagged lamin in these cells is predicted to result in nascent lamin meshwork
assembly, allowing visualization of the succession of lamin assembly using fluorescence and electron
microscopy. By comparing assembly of fluorescently tagged lamin mutants to assembly of wild type lamins, the
research will determine whether the lamin assembly pathway is altered by disease-causing lamin mutations.
 The research proposed will be conducted in the laboratory of Dr. Yixian Zheng at the Carnegie Institution
for Science Department of Embryology. Research will be carried out independently with biweekly guidance
provided by Dr. Zheng. Experimental training, along with trainin...

## Key facts

- **NIH application ID:** 10377350
- **Project number:** 5F32GM142145-02
- **Recipient organization:** CARNEGIE INSTITUTION OF WASHINGTON, D.C.
- **Principal Investigator:** Ross T Pedersen
- **Activity code:** F32 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $67,582
- **Award type:** 5
- **Project period:** 2021-04-01 → 2024-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10377350

## Citation

> US National Institutes of Health, RePORTER application 10377350, Molecular analysis of nuclear lamin assembly (5F32GM142145-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10377350. Licensed CC0.

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