Naslov
NDPK A and NDPK B – subcellular dynamics and interactions

Autori
Radić, Martina ; Šoštar, Marko ; Ćetković, Helena ; Weber, Igor ; Slade, Neda ; Herak Bosnar, Maja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
13th Multinational Congress on Microscopy : Book of Abstracts / Gajović, Andreja ; Weber, Igor ; Kovačević, Goran ; Čadež, Vida ; Šegota, Suzana ; Peharec Štefanić, Petra ; Vidoš, Ana - Zagreb : Institut Ruđer Bošković ; Hrvatsko mikroskopijsko društvo, 2017, 199-199

ISBN
978-953-7941-19-2

Skup
13th Multinational Congress on Microscopy

Mjesto i datum
Rovinj, Hrvatska, 24.09.2017. - 29.09.2017

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
NDPK A ; NDPK B ; live cell imaging ; FRET/FLIM technique

Sažetak
The Nme gene/protein family was named after the first member identified, Nm23-H1/Nme1, which is responsible for metastasis suppression in many tumor types. The Nme1/NDPKA and Nme2/NDPKB are two units of a well-known enzyme nucleoside- diphosphate kinase (NDPK), which transfers the terminal phosphate form (d)NTPs to (d)NDPs and is, therefore, responsible for the maintenance of the cellular nucleotide pool. Besides this housekeeping role, the Nme/NDPK A and B proteins have been assigned several additional biochemical functions such as the His-protein kinase and the scaffold function. There has also been evidence that they may function as transcriptional factors and as exonucleases. The two subunits can assemble into enzymatically active hexamers in all possible combinations (A6, A5B…..B6). Our hypothesis is that the isoenzyme assembly, with potentially distinct roles of the two subunits, differs between specific cellular physiological processes and locations. To be able to determine the potential diverse functions of the A and B subunits, as well as their spatially specific assembly, we subcloned their cDNAs into pEGFP and pmCherry fluorescent reporter vectors, and transfected human tumor cells in culture. By using live cell imaging techniques, we monitored their cellular dynamics with and without induction of DNA damage. Further, we investigated localization of the interaction between subunits by FRET/FLIM technique. Our preliminary results suggest that the assembly of heterohexamers is generally favorable compared to homohexamers. Additional experiments are needed to determine possible spatial specificity of these interactions.

Izvorni jezik
Engleski

Znanstvena područja
Biologija, Temeljne medicinske znanosti

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