The intracellular mobility of nuclear import receptors and NLS cargoes.

Abstract:

:We have investigated classical nuclear localization sequence (NLS) mediated protein trafficking by measuring biomolecular dynamics within living cells using two-photon fluorescence correlation spectroscopy. By directly observing the behavior of specific molecules in their native cellular environment, it is possible to uncover functional details that are not apparent from traditional biochemical investigations or functional assays. We show that the intracellular mobility of NLS cargoes and their import receptor proteins, karyopherin-alpha and karyopherin-beta, can be robustly measured and that quantitative comparison of intracellular diffusion coefficients provides new insights into nuclear transport mechanisms. Import cargo complexes are assembled throughout the cytoplasm, and their diffusion is slower than predicted by molecular weight due to specific interactions. Analysis of NLS cargo diffusion in the cytoplasm indicates that these interactions are likely disrupted by NLS cargo binding. Our results suggest that delivery of import receptors and NLS cargoes to nuclear pores may complement selective translocation through the pores as a functional mechanism for regulating transport of proteins into the nucleus.

journal_name

Biophys J

journal_title

Biophysical journal

authors

Wu J,Corbett AH,Berland KM

doi

10.1016/j.bpj.2009.01.050

subject

Has Abstract

pub_date

2009-05-06 00:00:00

pages

3840-9

issue

9

eissn

0006-3495

issn

1542-0086

pii

S0006-3495(09)00583-9

journal_volume

96

pub_type

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