Abstract
DSS1 is a small protein, highly conserved across different species. As a member of the intrinsically disordered protein family, DSS1 interacts with different protein partners, thus forming complexes involved in diverse biological mechanisms: DNA repair, regulation of protein homeostasis, mRNA export, etc. Additionally, DSS1 has a novel intriguing role in the post-translational protein modification named DSSylation. Oxidatively damaged proteins are targeted for removal with DSS1 and then degraded by proteasome. Yet, DSS1 involvement in the maintenance of genome integrity through homologous recombination is the only function well studied in Arabidopsis research. The fact that animal DSS1 shows wide multifunctionality imposes a need to investigate the additional roles of two Arabidopsis thaliana DSS1 homologs. Having in mind the universality of various biological processes, we considered the possibility of plant DSS1 involvement in cellular homeostasis maintenance during stress exposure. Using real-time PCR and immunoblot analysis, we investigated the profiles of DSS1 gene and protein expression under oxidative stress. We grew and selected the homozygous Arabidopsis mutant line, carrying the T-DNA intron insertion in the DSS1(V) gene. The mutant line was phenotypically described during plant development, and its sensitivity to oxidative stress was characterized. This is the first report which indicates that plant DSS1 gene expression has an altered profile under the influence of oxidative stress. dss1(V)−/− plants showed an increased sensitivity to oxidative stress, germinated faster than WT, but generally showed developmental delay in further stages. Our results indicate that the DSS1 protein could be a crucial player in the molecular mechanisms underlying plant abiotic stress responses.
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Data availability
The data and material that support the findings of this study are available from the corresponding author upon reasonable request.
Abbreviations
- ABA:
-
Abscisic acid
- BRCA2:
-
Breast cancer 2
- BSA:
-
Bovine serum albumin
- DNP hydrazine:
-
2,4-Dinitrophenilhydrazone
- DSS1:
-
Deleted in Split Hand/Split Foot Protein 1
- DTT:
-
Dithiothreitol
- EDTA:
-
Ethylenediaminetetraacetic acid
- FW:
-
Fresh weight
- HRP:
-
Horseradish peroxidase
- MDA:
-
Malondialdehyde
- MeJA:
-
Methyl jasmonate
- MV:
-
Methyl viologen
- NPC:
-
Nuclear pore complex
- PBST:
-
Phosphate buffered saline with Tween® 20
- PMSF:
-
Phenylmethylsulfonyl fluoride
- PSI:
-
Photosystem I
- PVDF:
-
Polyvinylidene difluoride membrane
- ROS:
-
Reactive oxygen species
- RPA:
-
Replication protein A
- SA:
-
Salicylic acid
- SDS:
-
Sodium dodecyl sulfate
- SEM1:
-
Suppressor of Exocyst Mutations 1
- SHSF:
-
Split hand/split foot malformation
- TBA:
-
Thiobarbituric acid
- TCA:
-
Trichloroacetic acid
- TREX-2:
-
Transcription-export-2
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Acknowledgments
The authors are grateful to Dr. Milorad Kojić for giving us an idea and immense motivation to conduct this investigation. We owe a debt of gratitude to Dr. Vesna Maksimović for encouragement and useful suggestions. Also, this project could not be accomplished without the support and technical advices of Dr. Jelena Brkljačić. We greatly acknowledge the ABRC and NASC for SALK_069888 seed stock distribution. We thank the Salk Institute Genomic Analysis Laboratory for providing the sequence-indexed Arabidopsis. This work was funded by the Ministry of Education, Science and Technological Development of the Republic of Serbia, registration number: 451-03-68/2020-14/200042 agreement on the implementation and financing of research work in 2020.
Funding
This work was funded by the Ministry of Education, Science and Technological Development of the Republic of Serbia, registration number: 451-03-68/2020-14/200042 agreement on the implementation and financing of research work in 2020.
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Gordana Timotijević and Ivana Nikolić contributed to the idea and experimental design and carried out experimental work. Data analysis and the first draft of the manuscript was written by Gordana Timotijević. Jelena Samardžić and Sofija Nešić contributed to experiments regarding the plant genotyping and provided critical feedback and analysis of the manuscript. All authors read and approved the final manuscript.
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Nikolić, I.P., Nešić, S.B., Samardžić, J.T. et al. Intrinsically disordered protein AtDSS1(V) participates in plant defense response to oxidative stress. Protoplasma 258, 779–792 (2021). https://doi.org/10.1007/s00709-020-01598-7
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DOI: https://doi.org/10.1007/s00709-020-01598-7