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Article

Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif

by 1,2,†, 3,4,5,†, 1,2 and 1,2,*
1
Asan Institute for Life Science, Asan Medical Center, Seoul 05505, Korea
2
Department of Convergence Medicine, University of Ulsan College of Medicine, Seoul 05505, Korea
3
Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul 03082, Korea
4
Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03082, Korea
5
Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul 03082, Korea
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Alfonso Baldi
Int. J. Mol. Sci. 2021, 22(15), 8293; https://doi.org/10.3390/ijms22158293
Received: 22 April 2021 / Revised: 24 July 2021 / Accepted: 29 July 2021 / Published: 2 August 2021
In vertebrates, nucleostemin (NS) is an important marker of proliferation in several types of stem and cancer cells, and it can also interact with the tumor-suppressing transcription factor p53. In the present study, the intra-nuclear diffusional dynamics of native NS tagged with GFP and two GFP-tagged NS mutants with deleted guanosine triphosphate (GTP)-binding domains were analyzed by fluorescence correlation spectroscopy. Free and slow binding diffusion coefficients were evaluated, either under normal culture conditions or under treatment with specific cellular proliferation inhibitors actinomycin D (ActD), 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB), or trichostatin A (TSA). When treated with ActD, the fractional ratio of the slow diffusion was significantly decreased in the nucleoplasm. The decrease was proportional to ActD treatment duration. In contrast, DRB or TSA treatment did not affect NS diffusion. Interestingly, it was also found that the rate of diffusion of two NS mutants increased significantly even under normal conditions. These results suggest that the mobility of NS in the nucleoplasm is related to the initiation of DNA or RNA replication, and that the GTP-binding motif is also related to the large change of mobility. View Full-Text
Keywords: fluorescence correlation spectroscopy; diffusion coefficient; nucleostemin; nuclear diffusion; ActD; DRB; TSA fluorescence correlation spectroscopy; diffusion coefficient; nucleostemin; nuclear diffusion; ActD; DRB; TSA
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MDPI and ACS Style

Pack, C.-G.; Jung, K.; Paulson, B.; Kim, J.K. Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif. Int. J. Mol. Sci. 2021, 22, 8293. https://doi.org/10.3390/ijms22158293

AMA Style

Pack C-G, Jung K, Paulson B, Kim JK. Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif. International Journal of Molecular Sciences. 2021; 22(15):8293. https://doi.org/10.3390/ijms22158293

Chicago/Turabian Style

Pack, Chan-Gi, Keehoon Jung, Bjorn Paulson, and Jun K. Kim. 2021. "Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif" International Journal of Molecular Sciences 22, no. 15: 8293. https://doi.org/10.3390/ijms22158293

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