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The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage

Radić, Martina; Šoštar, Marko; Weber, Igor; Ćetković, Helena; Slade, Neda; Herak Bosnar, Maja (2020) The Subcellular Localization and Oligomerization Preferences of NME1/NME2 upon Radiation-Induced DNA Damage. International journal of molecular sciences, 21 (7). ISSN 1422-0067

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Abstract

Nucleoside diphosphate kinases (NDPK/NME/Nm23) are enzymes composed of subunits NME1/NDPK A and NME2/NDPK B, responsible for the maintenance of the cellular (d)NTP pool and involved in other cellular processes, such as metastasis suppression and DNA damage repair. Although eukaryotic NDPKs are active only as hexamers, it is unclear whether other NME functions require the hexameric form, and how the isoenzyme composition varies in different cellular compartments. To examine the effect of DNA damage on intracellular localization of NME1 and NME2 and the composition of NME oligomers in the nucleus and the cytoplasm, we used live-cell imaging and the FRET/FLIM technique. We showed that exogenous NME1 and NME2 proteins co-localize in the cytoplasm of non-irradiated cells, and move simultaneously to the nucleus after gamma irradiation. The FRET/FLIM experiments imply that, after DNA damage, there is a slight shift in the homomer/heteromer balance between the nucleus and the cytoplasm. Collectively, our results indicate that, after irradiation, NME1 and NME2 engage in mutual functions in the nucleus, possibly performing specific functions in their homomeric states. Finally, we demonstrated that fluorophores fused to the N- termini of NME polypeptides produce the largest FRET effect and thus recommend this orientation for use in similar studies.

Item Type: Article
Uncontrolled Keywords: NME ; NDPK ; Nm23 ; nucleoside diphosphate kinase ; FRET ; FLIM ; live-cell imaging
Subjects: NATURAL SCIENCES > Biology
Divisions: Division of Molecular Biology
Division of Molecular Medicine
Projects:
Project titleProject leaderProject codeProject type
Otkrivanje novih proteinskih interakcija kao podloga za nove pristupe liječenju melanoma čovjekaSlade, NedaIP-2013-11-1615HRZZ
Struktura, funkcija i evolucija proteina Nme6/Nm23-H6Herak Bosnar, MajaIP-2016-06-4021HRZZ
Depositing User: Neda Slade
Date Deposited: 11 Dec 2020 09:19
Last Modified: 11 Dec 2020 09:19
URI: http://fulir.irb.hr/id/eprint/6126
DOI: 10.3390/ijms21072363

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