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Published on: June 9, 2023
Oxidant-induced cell-cycle delay in Saccharomyces cerevisiae: the involvement of the SWI6 transcription factor
Chii Shyang Fong1, Mark D Temple, Nazif Alic
1Ramaciotti Centre for Gene Function Analysis and School of Biotechnology and Biomolecular Sciences, UNSW, NSW, Australia.
Abstract:
Cells treated with low doses of linoleic acid hydroperoxide (LoaOOH) exhibit a cell-cycle delay that may provide a mechanism to overcome oxidative stress. Strains sensitive to LoaOOH from the genome-wide deletion collection were screened to identify deletants in which the cell-cycle delay phenotype was reduced. Forty-seven deletants were identified that were unable to mount the normal delay response, implicating the product of the deleted gene in the oxidant-mediated cell-cycle delay of the wild-type. Of these genes, SWI6 was of particular interest due to its role in cell-cycle progression through Start. The swi6 deletant strain was delayed on entry into the cell cycle in the absence of an oxidant, and oxidant addition caused no further delay. Transforming the swi6 deletant with SWI6 on a plasmid restored the G1 arrest in response to LoaOOH, indicating that Swi6p is involved in oxidant sensing leading to cell division delay. Micro-array studies identified genes whose expression in response to LoaOOH depended on SWI6. The screening identified 77 genes that were upregulated in the wild-type strain and concurrently downregulated in the swi6 deletant treated with LoaOOH. These data show that functions such as heat shock response, and glucose transport are involved in the response.
Insights
Low doses of linoleic acid hydroperoxide (LoaOOH) cause cell-cycle delay. The SWI6 gene is crucial for this oxidant-induced delay, impacting cellular responses to oxidative stress.
Area of Science:
- Cell Biology
- Molecular Biology
- Genetics
Background:
- Oxidative stress can trigger cellular defense mechanisms.
- Cell-cycle delay is a known response to environmental stressors.
- Identifying genes involved in stress response pathways is critical for understanding cellular resilience.
Purpose of the Study:
- To identify genes involved in the cell-cycle delay response to linoleic acid hydroperoxide (LoaOOH).
- To elucidate the role of SWI6 in oxidant-mediated cell-cycle arrest.
- To understand the downstream gene expression changes dependent on SWI6 during oxidative stress.
Main Methods:
- Genome-wide screening of deletion strains sensitive to LoaOOH.
- Phenotypic analysis of cell-cycle progression in wild-type and mutant strains.
- Plasmid-based complementation of the swi6 deletant.
- Micro-array analysis to assess gene expression changes.
Main Results:
- Forty-seven deletion strains showed a reduced cell-cycle delay in response to LoaOOH.
- The SWI6 gene was identified as critical for this delay.
- Loss of SWI6 resulted in a constitutive cell-cycle delay, unaffected by oxidant addition.
- SWI6 is required for the proper induction of genes involved in heat shock response and glucose transport upon LoaOOH treatment.
Conclusions:
- Swi6p plays a key role in sensing oxidants and mediating cell-cycle delay.
- SWI6 regulates a network of genes, including those involved in heat shock and glucose transport, as part of the oxidative stress response.
- This study reveals a novel function for SWI6 in cellular defense against oxidative damage.
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