Browsing by Author "Hema Rajaram"

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A single gene all3940 (Dps) overexpression in Anabaena sp. PCC 7120 confers multiple abiotic stress tolerance via proteomic alterations
(Springer Verlag, 2016) Om Prakash Narayan; Nidhi Kumari; Poonam Bhargava; Hema Rajaram; Lal Chand Rai
DNA-binding proteins (Dps) induced during starvation play an important role in gene regulation and maintaining homeostasis in bacteria. The nitrogen-fixing cyanobacterium, Anabaena PCC7120, has four genes annotated as coding for Dps; however, the information on their physiological roles is limiting. One of the genes coding for Dps, ‘all3940’ was found to be induced under different abiotic stresses in Anabaena and upon overexpression enhanced the tolerance of Anabaena to a multitude of stresses, which included salinity, heat, heavy metals, pesticide, and nutrient starvation. On the other hand, mutation in the gene resulted in decreased growth of Anabaena. The modulation in the levels of All3940 in Anabaena, achieved either by overexpression of the protein or mutation of the gene, resulted in changes in the proteome, which correlated well with the physiological changes observed. Proteins required for varied physiological activities, such as photosynthesis, carbon-metabolism, oxidative stress alleviation, exhibited change in protein profile upon modulation of All3940 levels in Anabaena. This suggested a direct or an indirect effect of All3940 on the expression of the above stress-responsive proteins, thereby enhancing tolerance in Anabaena PCC7120. Thus, All3940, though categorized as a Dps, is possibly a general stress protein having a global role in regulating tolerance to multitude of stresses in Anabaena. © 2015, Springer-Verlag Berlin Heidelberg.
Cd-induced cytosolic proteome changes in the cyanobacterium Anabaena sp. PCC7120 are mediated by LexA as one of the regulatory proteins
(Elsevier B.V., 2023) Akanksha Srivastava; Arvind Kumar; Subhankar Biswas; Vaibhav Srivastava; Hema Rajaram; Yogesh Mishra
LexA, a well-characterized transcriptional repressor of SOS genes in heterotrophic bacteria, has been shown to regulate diverse genes in cyanobacteria. An earlier study showed that LexA overexpression in a cyanobacterium, Anabaena sp. PCC7120 reduces its tolerance to Cd stress. This was later shown to be due to modulation of photosynthetic redox poising by LexA under Cd stress. However, due to the global regulatory nature of LexA and the prior prediction of AnLexA-box in a few heavy metal-responsive genes, we speculated that LexA has a broad role in Cd tolerance, with regulation over a variety of Cd stress-responsive genes in addition to photosynthetic genes. Thus, to further expand the knowledge on the regulatory role of LexA in Cd stress tolerance, a cytosolic proteome profiling of Anabaena constitutively overexpressing LexA upon Cd stress was performed. The proteomic study revealed 25 differentially accumulated proteins (DAPs) in response to the combined effect of LexA overexpression and Cd stress, and the other 11 DAPs exclusively in response to either LexA overexpression or Cd stress. The 36 identified proteins were related with a variety of functions, including photosynthesis, C-metabolism, antioxidants, protein turnover, post-transcriptional modifications, and a few unknown and hypothetical proteins. The regulation of LexA on corresponding genes, and six previously reported Cd efflux transporters, was further validated by the presence of AnLexA-boxes, transcript, and/or promoter analyses. In a nutshell, this study identifies the regulation of Anabaena LexA on several Cd stress-responsive genes of various functions, hence expanding the regulatory role of LexA under Cd stress. © 2023 Elsevier B.V.
Gamma (γ)-radiation stress response of the cyanobacterium Anabaena sp. PCC7120: Regulatory role of LexA and photophysiological changes
(Elsevier Ireland Ltd, 2023) Akanksha Srivastava; Arvind Kumar; Subhankar Biswas; Rajender Kumar; Vaibhav Srivastava; Hema Rajaram; Yogesh Mishra
High radioresistance of the cyanobacterium, Anabaena sp. PCC7120 has been attributed to efficient DNA repair, protein recycling, and oxidative stress management. However, the regulatory network involved in these batteries of responses remains unexplored. In the present study, the role of a global regulator, LexA in modulating gamma (γ)-radiation stress response of Anabaena was investigated. Comparison of the cytosolic proteome profiles upon γ-radiation in recombinant Anabaena strains, AnpAM (vector-control) and AnlexA+ (LexA-overexpressing), revealed 41 differentially accumulated proteins, corresponding to 29 distinct proteins. LexA was found to be involved in the regulation of 27 of the corresponding genes based on the presence of AnLexA-Box, EMSA, and/or qRT-PCR studies. The majority of the regulated genes were found to be involved in C-assimilation either through photosynthesis or C-catabolism and oxidative stress alleviation. Photosynthesis, measured in terms of PSII photophysiological parameters and thylakoid membrane proteome was found to be affected by γ-radiation in both AnpAM and AnlexA+ cells, with LexA affecting them even under control growth conditions. Thus, LexA functioned as one of the transcriptional regulators involved in modulating γ-radiation stress response in Anabaena. This study could pave the way for a deeper understanding of the regulation of γ-radiation-responsive genes in cyanobacteria at large. © 2022 Elsevier B.V.
Physiological and thylakoid proteome analyses of Anabaena sp. PCC 7120 for monitoring the photosynthetic responses under cadmium stress
(Elsevier B.V., 2021) Akanksha Srivastava; Subhankar Biswas; Sandhya Yadav; Arvind Kumar; Hema Rajaram; Vaibhav Srivastava; Yogesh Mishra
Photosynthetic organisms are highly susceptible to cadmium (Cd) as it interferes with the structural and functional aspects of the photosynthesis. As a major group of photosynthetic prokaryotes, cyanobacteria are also affected by Cd. The inhibitory effects of Cd on cyanobacterial photosynthesis have been studied from an early stage, but the mechanism of Cd toxicity is still unclear. Therefore, we investigated the photosynthetic responses of Anabaena sp. PCC 7120 under Cd stress (LC50) at physiological and thylakoid proteome levels via chlorophyll a fluorescence measurements and blue native (BN)-SDS PAGE, respectively. The findings revealed that Cd exposure triggered the intracellular ROS production and negatively affected the photosynthetic performance. With Cd exposure, effective photochemical quantum yield of PSII (Y(II)) and photochemical quenching efficiency (qP and qL) were significantly reduced, whereas the non-regulated energy dissipation (Y(NO)) was increased. Fast fluorescence kinetic measurements showed that Cd exposure resulted in the inactivation of PSII reaction centres, which interfered with the transfer of energy from antenna complexes to reaction centre, decreased the ability to convert excitation energy into electron transport, increased thermal dissipation, and thus decreased linear electron flow efficiency. Conversely, the efficiency of electron transfer from intermediate carriers to final PSI acceptors was increased, which might be related to the induction of PSI-based cyclic electron flow. This assumption was confirmed by an enhanced post-illumination fluorescence transient. Consistent with these observations, BN-SDS PAGE showed that Cd exposure significantly decreased PSII subunit proteins accumulation, but slightly increased PSI proteins accumulation. Additionally, only minor reductions in the accumulation of ATP synthase/NDH-1 and Cytochrome b6f subunit proteins were observed. The transcript levels of most of the selected subunit proteins were in accordance with BN-SDS PAGE. Collectively, our experiments show that a shift to PSI-based cyclic electron flow was a protective strategy of Anabaena sp. PCC 7120 against Cd-induced PSII photo-damage. © 2021 Elsevier B.V.
Regulatory role of LexA in modulating photosynthetic redox poise and cadmium stress tolerance in the cyanobacterium, Anabaena sp. PCC7120
(Elsevier B.V., 2022) Akanksha Srivastava; Arvind Kumar; Subhankar Biswas; Vaibhav Srivastava; Hema Rajaram; Yogesh Mishra
Strategies developed by organisms to overcome disruption in redox poise of photosynthetic electron transport chain (pETC) are important for its survival under abiotic stress. The process needs to be tightly regulated for optimal functioning. While the redox poising processes are well known in cyanobacteria, understanding of their regulatory network is lacking. Since LexA is one of the known global regulators of stress response in the cyanobacterium Anabaena sp. PCC7120, its role in pETC redox poising was investigated using cadmium (Cd) as an abiotic stressor to disrupt photosynthesis. Assessment of the photosynthetic responses of recombinant Anabaena strains, AnlexA+ (LexA-overexpressing) and AnpAM (vector control), under unstressed and Cd-stressed conditions using transmission electron microscopy (TEM) and chlorophyll a fluorescence, indicated that some pETC redox poising responses, including PSII photodamage, energy dissipation, PSI photoprotection, and NDH-mediated cyclic electron flow were decreased in AnlexA+ under unstressed conditions. Disturbance in pETC redox poise during Cd stress observed in Anabaena was accentuated upon overexpression of LexA. The decreased photodamage of PSII and increased photoinhibition of PSI in AnlexA+ in the presence or absence of Cd stress, correlated well with the changes in pETC complexes observed in blue native (BN)-PAGE and the regulation of over 70 of the 90 pETC component genes by LexA demonstrated through transcript, electromobility shift assay (EMSA), and bioinformatics studies. In a nutshell, LexA has been identified as one of the regulators involved in the streamlining of pETC redox poising responses under normal growth and during abiotic stress through transcriptional regulation of some of the redox-controlled pETC component genes. © 2022 Elsevier B.V.