DOI: 10.1242/jcs.114.24.4621. Regulation of functional nuclear pore size in fibroblasts

Regulation of functional nuclear pore size in fibroblasts

10.1242/jcs.114.24.4621

Crossref journal-article

The Company of Biologists

Journal of Cell Science (237)

Abstract

Protein-NLS-coated gold particles up to approximately 250 Å in diameter are transported through the nuclear pores in normal, proliferating BALB/c 3T3 cells. This size can increase or decrease, depending on cellular activity. It has been suggested that increases in functional pore size are related to a reduction in the amount of available p53. To further test this hypothesis, we investigated the effects of cycloheximide and pifithrin-α, which inhibits p53-dependent transcriptional activation, on nuclear transport. After 3 hours in cycloheximide, there was a significant increase in the size of the gold particles that entered the nucleoplasm. When the incubation period was extended to 6 hours or longer, transport capacity returned to the control level. By using proteasome inhibitors, it was shown that the cycloheximide-dependent increase in functional pore size was due to the inhibition of protein synthesis, consistent with the fact that p53 is a short-lived protein, and requires the activity of at least two different factors. Although cycloheximide increases the functional diameter of the channel available for signal-mediated transport by approximately 60 Å, it had no significant effect on either the import rate of small NLS-containing substrates (FITC-BSA-NLS), or passive diffusion of fluorescent-labeled proteins across the envelope. This suggests that changes in transport capacity were not caused by an increase in overall pore diameter but instead are due to a transient increase in pore size that accompanies signal-mediated transport. Pifithrin-α also caused an increase in functional pore diameter without altering the import rate of FITC-BSA-NLS, providing further support for the view that p53 can initiate changes in nuclear transport capacity.

Bibliography

Feldherr, C. M., Akin, D., & Cohen, R. J. (2001). Regulation of functional nuclear pore size in fibroblasts. Journal of Cell Science, 114(24), 4621â4627.

Authors 3

Carl M. Feldherr (first)
Debra Akin (additional)
Robert J. Cohen (additional)

References 27 Referenced 36

Dates

Type	When
Created	4 years, 4 months ago (April 25, 2021, 8:34 p.m.)
Deposited	4 years, 4 months ago (April 25, 2021, 8:34 p.m.)
Indexed	4 months, 4 weeks ago (March 30, 2025, 5:44 p.m.)
Issued	23 years, 8 months ago (Dec. 15, 2001)
Published	23 years, 8 months ago (Dec. 15, 2001)
Published Print	23 years, 8 months ago (Dec. 15, 2001)

Funders 0

None

BibTeX

@article{Feldherr_2001, title={Regulation of functional nuclear pore size in fibroblasts}, volume={114}, ISSN={0021-9533}, url={http://dx.doi.org/10.1242/jcs.114.24.4621}, DOI={10.1242/jcs.114.24.4621}, number={24}, journal={Journal of Cell Science}, publisher={The Company of Biologists}, author={Feldherr, Carl M. and Akin, Debra and Cohen, Robert J.}, year={2001}, month=dec, pages={4621–4627} }

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  "abstract": "<jats:p>Protein-NLS-coated gold particles up to approximately 250 A\u030a in diameter are transported through the nuclear pores in normal, proliferating BALB/c 3T3 cells. This size can increase or decrease, depending on cellular activity. It has been suggested that increases in functional pore size are related to a reduction in the amount of available p53. To further test this hypothesis, we investigated the effects of cycloheximide and pifithrin-\u03b1, which inhibits p53-dependent transcriptional activation, on nuclear transport. After 3 hours in cycloheximide, there was a significant increase in the size of the gold particles that entered the nucleoplasm. When the incubation period was extended to 6 hours or longer, transport capacity returned to the control level. By using proteasome inhibitors, it was shown that the cycloheximide-dependent increase in functional pore size was due to the inhibition of protein synthesis, consistent with the fact that p53 is a short-lived protein, and requires the activity of at least two different factors. Although cycloheximide increases the functional diameter of the channel available for signal-mediated transport by approximately 60 A\u030a, it had no significant effect on either the import rate of small NLS-containing substrates (FITC-BSA-NLS), or passive diffusion of fluorescent-labeled proteins across the envelope. This suggests that changes in transport capacity were not caused by an increase in overall pore diameter but instead are due to a transient increase in pore size that accompanies signal-mediated transport. Pifithrin-\u03b1 also caused an increase in functional pore diameter without altering the import rate of FITC-BSA-NLS, providing further support for the view that p53 can initiate changes in nuclear transport capacity.</jats:p>",
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