Albany 2019: 20th Conversation - Abstracts

category image Albany 2019
Conversation 20
June 11-15 2019
Adenine Press (2019)

Applying quantitative single molecule localization microscopy to probe the mechanism of nucleocytoplasmic transport

Transport between the nucleus and cytoplasm is tightly regulated by nuclear pore complexes (NPCs). While these intricate molecular machines obstruct passage of non-specific biomolecules, cargo-laden transport proteins are efficiently transported across the nuclear membrane. NPCs are essential for transcription, signaling, and other fundamental cellular processes. As such, errors in the nucleocytoplasmic transport can cause cellular dysregulation. Dysregulated transport has been observed in both neurodegenerative disorders and cancer. Despite its critical role in health and disease, specific mechanism(s) of nucleocytoplasmic transport currently remain unresolved. Understanding the NPC transport mechanism(s) has been hindered by the 1) complexity of NPC geometry, 2) disordered nature of barrier elements called FG-Nups, and 3) challenge of measuring millisecond transport events at nanometer scales in the intricate cellular environment. To probe potential transport mechanisms, we combined biochemical approaches with advanced quantitative single molecule localization microcopy (Golfetto et al., 2018). We measured both binding and diffusion of transport proteins on oriented FG-Nup monolayers. Our ultimate goal is to obtain critical information about both physiological and pathological NPC transport at the single molecule level.

This research has been supported by STOP Cancer Foundation and Beckman Research Institute.


Golfetto O., Wakefield D.L., Cacao E.E., Avery K., Kenyon V., Jorand R., Tobin S.J., Biswas S., Gutierrez J., Clinton R., Ma Y., Horne D.A., Williams J.C., Jovanovic-Talisman, T. (2018). A Platform to Enhance Quantitative Single Molecule Localization Microscopy. J. Am. Chem. Soc., 140, 12785.

Kathleen M. Lennon
Devin L. Wakefield
Matthew S. Brehove
Ottavia Golfetto
Sunetra Biswas
Tijana Jovanovic-Talisman

Department of Molecular Medicine
Beckman Research Institute
City of Hope
Duarte, CA 91010

Ph: (626) 218-9067
E-mail: ttalisman@coh.org