Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest
Mitochondria are semi-autonomous organelles containing their own DNA (mtDNA), which is replicated independently of nuclear DNA (nDNA). While cell cycle arrest halts nDNA replication, mtDNA replication continues. In Saccharomyces cerevisiae, flow cytometry enables semi-quantitative estimation of mtDN...
Saved in:
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2024-12-01
|
| Series: | Frontiers in Cell and Developmental Biology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2024.1497652/full |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1846121049906741248 |
|---|---|
| author | Elena Yu Potapenko Nataliia D. Kashko Dmitry A. Knorre |
| author_facet | Elena Yu Potapenko Nataliia D. Kashko Dmitry A. Knorre |
| author_sort | Elena Yu Potapenko |
| collection | DOAJ |
| description | Mitochondria are semi-autonomous organelles containing their own DNA (mtDNA), which is replicated independently of nuclear DNA (nDNA). While cell cycle arrest halts nDNA replication, mtDNA replication continues. In Saccharomyces cerevisiae, flow cytometry enables semi-quantitative estimation of mtDNA levels by measuring the difference in signals between cells lacking mtDNA and those containing mtDNA. In this study, we used flow cytometry to investigate mtDNA accumulation in yeast cells under G1 and G2 phase cell cycle arrest conditions utilising thermosensitive mutants cdc4-3 and cdc15-2. In line with the previous studies, cell cycle arrest induced a several-fold accumulation of mtDNA in both mutants. The total DNA levels in arrested cells correlated with cell forward scattering, suggesting a relationship between individual cell mtDNA quantity and size. In cell cycle-arrested cells, we observed no correlation between cell size and intercellular mtDNA copy number variability. This implies that as cell size increases during arrest, the mtDNA content remains within a specific limited range for each size class. This observation suggests that mtDNA quantity control mechanisms can function in cell cycle-arrested cells. |
| format | Article |
| id | doaj-art-bb75e8b15bbb459186df5b898b92c5e3 |
| institution | Kabale University |
| issn | 2296-634X |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj-art-bb75e8b15bbb459186df5b898b92c5e32024-12-16T06:18:37ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2024-12-011210.3389/fcell.2024.14976521497652Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrestElena Yu Potapenko0Nataliia D. Kashko1Dmitry A. Knorre2A. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, RussiaFaculty of Bioengineering and Bioinformatics, Lomonosov Moscow State University, Moscow, RussiaA. N. Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Moscow, RussiaMitochondria are semi-autonomous organelles containing their own DNA (mtDNA), which is replicated independently of nuclear DNA (nDNA). While cell cycle arrest halts nDNA replication, mtDNA replication continues. In Saccharomyces cerevisiae, flow cytometry enables semi-quantitative estimation of mtDNA levels by measuring the difference in signals between cells lacking mtDNA and those containing mtDNA. In this study, we used flow cytometry to investigate mtDNA accumulation in yeast cells under G1 and G2 phase cell cycle arrest conditions utilising thermosensitive mutants cdc4-3 and cdc15-2. In line with the previous studies, cell cycle arrest induced a several-fold accumulation of mtDNA in both mutants. The total DNA levels in arrested cells correlated with cell forward scattering, suggesting a relationship between individual cell mtDNA quantity and size. In cell cycle-arrested cells, we observed no correlation between cell size and intercellular mtDNA copy number variability. This implies that as cell size increases during arrest, the mtDNA content remains within a specific limited range for each size class. This observation suggests that mtDNA quantity control mechanisms can function in cell cycle-arrested cells.https://www.frontiersin.org/articles/10.3389/fcell.2024.1497652/fullmtDNAyeastmtDNA copy number controlmtDNA copy numbercell cycle arrestcell cycle defect |
| spellingShingle | Elena Yu Potapenko Nataliia D. Kashko Dmitry A. Knorre Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest Frontiers in Cell and Developmental Biology mtDNA yeast mtDNA copy number control mtDNA copy number cell cycle arrest cell cycle defect |
| title | Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest |
| title_full | Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest |
| title_fullStr | Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest |
| title_full_unstemmed | Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest |
| title_short | Flow-cytometry reveals mitochondrial DNA accumulation in Saccharomyces cerevisiae cells during cell cycle arrest |
| title_sort | flow cytometry reveals mitochondrial dna accumulation in saccharomyces cerevisiae cells during cell cycle arrest |
| topic | mtDNA yeast mtDNA copy number control mtDNA copy number cell cycle arrest cell cycle defect |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2024.1497652/full |
| work_keys_str_mv | AT elenayupotapenko flowcytometryrevealsmitochondrialdnaaccumulationinsaccharomycescerevisiaecellsduringcellcyclearrest AT nataliiadkashko flowcytometryrevealsmitochondrialdnaaccumulationinsaccharomycescerevisiaecellsduringcellcyclearrest AT dmitryaknorre flowcytometryrevealsmitochondrialdnaaccumulationinsaccharomycescerevisiaecellsduringcellcyclearrest |