Cargando…

Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells

Thymidylate synthase (TS), dihydrofolate reductase (DHFR), and serine hydroxymethyltransferase (SHMT) constitute the thymidylate synthesis cycle providing thymidylate for DNA synthesis and repair. Our previous studies indicated that TS and DHFR are the substrates of protein kinase CK2. This work has...

Descripción completa

Detalles Bibliográficos
Autores principales: Wińska, Patrycja, Widło, Łukasz, Senkara, Elżbieta, Koronkiewicz, Mirosława, Cieśla, Jarosław M., Krzyśko, Alicja, Skierka, Katarzyna, Cieśla, Joanna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914513/
https://www.ncbi.nlm.nih.gov/pubmed/35281258
http://dx.doi.org/10.3389/fmolb.2022.847829
_version_ 1784667725793067008
author Wińska, Patrycja
Widło, Łukasz
Senkara, Elżbieta
Koronkiewicz, Mirosława
Cieśla, Jarosław M.
Krzyśko, Alicja
Skierka, Katarzyna
Cieśla, Joanna
author_facet Wińska, Patrycja
Widło, Łukasz
Senkara, Elżbieta
Koronkiewicz, Mirosława
Cieśla, Jarosław M.
Krzyśko, Alicja
Skierka, Katarzyna
Cieśla, Joanna
author_sort Wińska, Patrycja
collection PubMed
description Thymidylate synthase (TS), dihydrofolate reductase (DHFR), and serine hydroxymethyltransferase (SHMT) constitute the thymidylate synthesis cycle providing thymidylate for DNA synthesis and repair. Our previous studies indicated that TS and DHFR are the substrates of protein kinase CK2. This work has been aimed at the elucidation of the effect of CK2 activity on cell cycle progression, thymidylate synthesis enzyme expression and localization, and the role of CK2-mediated TS phosphorylation in in vitro di- and trimolecular complex formation. The results were obtained by means of western blot, confocal microscopy, flow cytometry, quantitative polymerase chain reaction (QPCR), quartz crystal microbalance with dissipation monitoring (QCM-D), and microthermophoresis (MST). Our research indicates that CK2 inhibition does not change the levels of the transcripts; however, it affects the protein levels of DHFR and TS in both tested cell lines, i.e., A549 and CCRF-CEM, and the level of SHMT1 in CCRF-CEM cells. Moreover, we show that CK2-mediated phosphorylation of TS enables the protein (pTS) interaction with SHMT1 and leads to the stability of the tri-complex containing SHMT1, DHFR, and pTS. Our results suggest an important regulatory role of CK2-mediated phosphorylation for inter- and intracellular protein level of enzymes involved in the thymidylate biosynthesis cycle.
format Online
Article
Text
id pubmed-8914513
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-89145132022-03-12 Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells Wińska, Patrycja Widło, Łukasz Senkara, Elżbieta Koronkiewicz, Mirosława Cieśla, Jarosław M. Krzyśko, Alicja Skierka, Katarzyna Cieśla, Joanna Front Mol Biosci Molecular Biosciences Thymidylate synthase (TS), dihydrofolate reductase (DHFR), and serine hydroxymethyltransferase (SHMT) constitute the thymidylate synthesis cycle providing thymidylate for DNA synthesis and repair. Our previous studies indicated that TS and DHFR are the substrates of protein kinase CK2. This work has been aimed at the elucidation of the effect of CK2 activity on cell cycle progression, thymidylate synthesis enzyme expression and localization, and the role of CK2-mediated TS phosphorylation in in vitro di- and trimolecular complex formation. The results were obtained by means of western blot, confocal microscopy, flow cytometry, quantitative polymerase chain reaction (QPCR), quartz crystal microbalance with dissipation monitoring (QCM-D), and microthermophoresis (MST). Our research indicates that CK2 inhibition does not change the levels of the transcripts; however, it affects the protein levels of DHFR and TS in both tested cell lines, i.e., A549 and CCRF-CEM, and the level of SHMT1 in CCRF-CEM cells. Moreover, we show that CK2-mediated phosphorylation of TS enables the protein (pTS) interaction with SHMT1 and leads to the stability of the tri-complex containing SHMT1, DHFR, and pTS. Our results suggest an important regulatory role of CK2-mediated phosphorylation for inter- and intracellular protein level of enzymes involved in the thymidylate biosynthesis cycle. Frontiers Media S.A. 2022-02-25 /pmc/articles/PMC8914513/ /pubmed/35281258 http://dx.doi.org/10.3389/fmolb.2022.847829 Text en Copyright © 2022 Wińska, Widło, Senkara, Koronkiewicz, Cieśla, Krzyśko, Skierka and Cieśla. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Molecular Biosciences
Wińska, Patrycja
Widło, Łukasz
Senkara, Elżbieta
Koronkiewicz, Mirosława
Cieśla, Jarosław M.
Krzyśko, Alicja
Skierka, Katarzyna
Cieśla, Joanna
Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells
title Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells
title_full Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells
title_fullStr Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells
title_full_unstemmed Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells
title_short Inhibition of Protein Kinase CK2 Affects Thymidylate Synthesis Cycle Enzyme Level and Distribution in Human Cancer Cells
title_sort inhibition of protein kinase ck2 affects thymidylate synthesis cycle enzyme level and distribution in human cancer cells
topic Molecular Biosciences
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8914513/
https://www.ncbi.nlm.nih.gov/pubmed/35281258
http://dx.doi.org/10.3389/fmolb.2022.847829
work_keys_str_mv AT winskapatrycja inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT widłołukasz inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT senkaraelzbieta inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT koronkiewiczmirosława inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT cieslajarosławm inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT krzyskoalicja inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT skierkakatarzyna inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells
AT cieslajoanna inhibitionofproteinkinaseck2affectsthymidylatesynthesiscycleenzymelevelanddistributioninhumancancercells