Dissertations, Theses, and Capstone Projects

Date of Degree

9-2026

Document Type

Doctoral Dissertation

Degree Name

Doctor of Philosophy

Program

Biology

Advisor

Hualin Zhong

Committee Members

Diego Loayza

Weigang Qiu

Songtao Jia

Frederick R. Cross

Subject Categories

Bioinformatics | Biology | Cell Biology | Molecular Biology | Molecular Genetics

Keywords

Nuclear pore complex, Nuclear basket, Nup211, Schizosaccharomyces pombe, mRNA export, Gene expression, Transcriptomics, Proximity labeling

Abstract

Nuclear pore complexes (NPCs) are evolutionarily conserved structures that span the nuclear envelope and serve as gateways between the nucleus and cytoplasm. On the nuclear face of NPCs, the nuclear basket is formed primarily by nucleoporins such as TPR and its orthologs. TPR family proteins play important roles in mRNA export and have also been implicated in diverse aspects of gene regulation, including chromatin organization and transcription. In the fission yeast Schizosaccharomyces pombe, the TPR ortholog Nup211 is essential for cell viability; however, the basis of this essentiality remains unclear. In this study, I used genetic, transcriptomic, and proteomic approaches to investigate the molecular functions of Nup211. To identify a minimal region of Nup211 that is sufficient for viability, constructs expressing a series of Nup211 fragments were introduced into a conditional depletion strain (nup211-so). These experiments demonstrated that the N-terminal 655 residues are sufficient to support viability. Using CRISPR/Cas9 genome editing, I generated a strain expressing Nup2111-655 from the endogenous locus and confirmed this finding. To determine the impact of Nup211 depletion on gene expression, I performed RNA sequencing. Data analyses revealed global changes in transcript abundance, with most differentially expressed transcripts being upregulated. Finally, I performed co-immunoprecipitation and TurboID proximity labeling to identify Nup211-interacting factors. In addition to other nuclear basket proteins, these approaches identified proteasome components and chromatin-associated factors, suggesting that Nup211 functions in coordination with the proteasome and chromatin. Together, these findings provide a framework for understanding the essential role of Nup211 and, more broadly, how nuclear basket proteins regulate gene expression at multiple levels.

This work is embargoed and will be available for download on Saturday, September 30, 2028

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