Heterologous Expression of Stress-Responsive DUF538 Domain Containing Protein and its Morpho-Biochemical Consequences

Abstract

As a usual response, plants induce/activate various proteins which are thought to be involved in defense mechanisms against the biotic and abiotic stresses they may be confronted with. The novel DUF538 domain containing proteins with unknown functions have been found to be induced/activated in response to different environmental stress stimuli in plants. In order to perform biochemical studies with these new plant stress-responsive proteins, a cDNA containing DUF538 domain was amplified from Celosia cristata full-length leaf expression library using a specific primer set. The isolated cDNA was subsequently expressed in Escherichia coli as a part of maltose-binding fusion protein (MBP-DUF538 construct) and purified at the yield of about 32 mg per liter of cell culture by affinity chromatography without affecting the recombinant bacterial cell growth. The purified fusion product was exogenously applied (10 μg per 4 cm(2)) on the leaves of Nicotiana tobaccum L. The results revealed that fused DUF538 protein does not induce morphological reposes, but elevates redox enzyme activities including catalase, peroxidase, polyphenol oxidase and phenyalanine ammonia lyase. This is the first time ever time report with respect to the heterologous expression of a plant stress-responsive DUF538 domain that may provide a basis to study its physiological roles and biochemical activities in vitro and in vivo.

A survey of genes involved in Arachis stenosperma resistance to Meloidogyne arenaria race 1

“…DUF protein was shown to elevate activity of enzymes that induce the HR response, such as catalase, peroxidase, polyphenol oxidase and phenyalanine ammonia lyase, when applied to tobacco leaves (Gholizadeh 2011). In this study, AsDUF was upregulated in the three time points studied, suggesting its role in the induction of these ROS enzymes.…”

“…Another protein family containing a specific domain related to plant defence is the DUF protein family, identified in Arabidopsis, rice and tomato plants grown under various environmental stress conditions, such as nutrient deficiency, Agrobacterium-induced crown gall, and mixed elicitors (Brunings et al 2009;Gholizadeh 2011). DUF protein was shown to elevate activity of enzymes that induce the HR response, such as catalase, peroxidase, polyphenol oxidase and phenyalanine ammonia lyase, when applied to tobacco leaves (Gholizadeh 2011).…”

DUF538 protein superfamily is predicted to be chlorophyll hydrolyzing enzymes in plants

“…On the basis of the high phosphorylation potential, DUF538 proteins have been predicted to play important regulatory roles in different stress-challenged plants (Nakagami et al 2010). Our report has been revealed that the exogenously applied fusion form of a DUF538 protein by using a plant tissue abrading material activates the redox system of the plant cells (Gholizadeh 2011).…”

“…Introduction DUF538 protein superfamily consists of several plant proteins of unknown functions. They have been distributed in wide ranges of monocotyledonous and dicotyledonous plant species (Gholizadeh 2011;Takahashi et al 2013). Their molecular weights are about 19-21 kDa, encoding about 170 amino acids.…”

Maltose-binding protein switches programmed cell death in Nicotiana glutinosa leaf cells

“…Most of the expressed proteins are usually able to fold into their biologically active conformations when fused to MBP [20][21][22]. The MBP-fused proteins are often used for the basic investigations including their morpho-molecular interactions with the eukaryotic cells/tissues and several biological assays without the cleavage of their fusion parts [23,24]. Therefore, it is essential to understood the likely interactive responses of eukaryotic cells with MBP.…”