Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif
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-tag...
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doaj-fe49c87b02d24076be6e4f1bb0a6d4f32021-08-06T15:26:11ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-08-01228293829310.3390/ijms22158293Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding MotifChan-Gi Pack0Keehoon Jung1Bjorn Paulson2Jun Ki Kim3Asan Institute for Life Science, Asan Medical Center, Seoul 05505, KoreaDepartment of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul 03082, KoreaAsan Institute for Life Science, Asan Medical Center, Seoul 05505, KoreaAsan Institute for Life Science, Asan Medical Center, Seoul 05505, KoreaIn 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.https://www.mdpi.com/1422-0067/22/15/8293fluorescence correlation spectroscopydiffusion coefficientnucleosteminnuclear diffusionActDDRB |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Chan-Gi Pack Keehoon Jung Bjorn Paulson Jun Ki Kim |
spellingShingle |
Chan-Gi Pack Keehoon Jung Bjorn Paulson Jun Ki Kim Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif International Journal of Molecular Sciences fluorescence correlation spectroscopy diffusion coefficient nucleostemin nuclear diffusion ActD DRB |
author_facet |
Chan-Gi Pack Keehoon Jung Bjorn Paulson Jun Ki Kim |
author_sort |
Chan-Gi Pack |
title |
Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif |
title_short |
Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif |
title_full |
Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif |
title_fullStr |
Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif |
title_full_unstemmed |
Mobility of Nucleostemin in Live Cells Is Specifically Related to Transcription Inhibition by Actinomycin D and GTP-Binding Motif |
title_sort |
mobility of nucleostemin in live cells is specifically related to transcription inhibition by actinomycin d and gtp-binding motif |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-08-01 |
description |
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. |
topic |
fluorescence correlation spectroscopy diffusion coefficient nucleostemin nuclear diffusion ActD DRB |
url |
https://www.mdpi.com/1422-0067/22/15/8293 |
work_keys_str_mv |
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