Abstract
WD40 domain-containing proteins are one of the eukaryotes’ most ancient and ubiquitous protein families. Little is known about the presence and function of these proteins in cyanobacteria in general and Anabaena in particular. In silico analysis confirmed the presence of WD40 repeats. Gene expression analysis indicated that the transcript levels of both the target proteins were up-regulated up to 4 fold in Cd and drought and 2–3 fold in heat, salt, and UV-B stress. Using a fluorescent oxidative stress indicator, we showed that the recombinant proteins were scavenging reactive oxygen species (ROS) (4–5 fold) more efficiently than empty vectors. Chromatin immunoprecipitation analysis (ChIP) and electrophoretic mobility shift assay (EMSA) revealed that the target proteins function as transcription factors after binding to the promoter sequences. The presence of kinase activity (2–4 fold) in the selected proteins indicated that these proteins could modulate the functions of other cellular proteins under stress conditions by inducing phosphorylation of specific amino acids. The chosen proteins also demonstrated interaction with Zn, Cd, and Cu (1.4–2.5 fold), which might stabilize the proteins’ structure and biophysical functions under multiple abiotic stresses. The functionally characterized Alr0671 and All2352 proteins act as transcription factors and offer tolerance to agriculturally relevant abiotic stresses.
Key message
Alr0671 and All2352 are novel WD40 proteins of Anabaena capable of regulating biochemical functions and abiotic stress tolerance by acting as a transcription factor and mediating DNA-protein interaction.
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Data Availability
All data supporting the present study’s findings are available within the paper and its supplementary files. All the software used for in silico analysis is publically available.
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Acknowledgements
L.C. Rai thanks the National Academy of Science, India for the NASI Senior Scientist Platinum Jubilee Fellowship, the Indian Council of Agricultural Research-National Bureau of Agriculturally Important Microorganisms (ICAR-NBAIM) for financial support; Dr. Krishna Kumar Rai is thankful to the National Academy of Science, India for RA ship. Dr. Shilpi Singh thanks CSIR for the Senior Research Associate award. Dr. Ruchi Rai is thankful to DST, New Delhi, for Women Scientist Scheme A (WOSA) award.
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KKR, RR, and LCR designed the experiments. KKR conducted the experiments and prepared the initial draft of the manuscript. SS performed in-silico analysis, and RR and LCR analyzed the data and critically reviewed the paper.
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Rai, K.K., Singh, S., Rai, R. et al. Functional characterization of two WD40 family proteins, Alr0671 and All2352, from Anabaena PCC 7120 and deciphering their role in abiotic stress management. Plant Mol Biol 110, 545–563 (2022). https://doi.org/10.1007/s11103-022-01306-4
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DOI: https://doi.org/10.1007/s11103-022-01306-4