Non-native ligands define the active site of Pennisetum glaucum (L.) R. Br dehydroascorbate reductase.

Dehydroascorbate reductase (DHAR), a member of the glutathione-S-transferase (GST) family, reduces dehydroascorbate (DHA) to ascorbate (AsA; Vitamin-C) in a glutathione (GSH)-dependent manner and in doing so, replenishes the critical AsA pool of the cell. To understand the enzyme mechanism in detail, we determined the crystal structure of a plant DHAR from Pennisetum glaucum (PgDHAR) using Iodide-Single Anomalous Dispersion (SAD) and Molecular replacement methods, in two different space groups. Here, we show PgDHAR in complex with two non-native ligands, viz.

Dengue NS3, an RNAi suppressor, modulates the human miRNA pathways through its interacting partner.

RNAi acts as a host immune response against non-self molecules, including viruses. Viruses evolved to neutralize this response by expressing suppressor proteins. In the present study, we investigated dengue virus non structural protein 3 (dvNS3), for its RNAi-suppressor activity in human cell lines.

Molecular and biochemical characterization of dehydroascorbate reductase from a stress adapted C4 plant, pearl millet [Pennisetum glaucum (L.) R. Br].

Key Message: PgDHAR was isolated from Pennisetum glaucum. PgDHAR responded to abiotic stress and exhibited enzyme activity at broad ranges of temperature, pH and substrate concentrations suggesting its role in stress tolerance.

Abstract: Dehydroascorbate reductase (EC 1.

Biochemical and molecular analyses of copper-zinc superoxide dismutase from a C4 plant Pennisetum glaucum reveals an adaptive role in response to oxidative stress.

Superoxide dismutases (SODs) form the foremost line of defense against ROS in aerobes. Pennisetum glaucum cDNA library is constructed to isolate superoxide dismutase cDNA clone (PgCuZnSOD) of 798 bp comprising 5'UTR (111 bp), an ORF (459 bp) and 3'UTR (228 bp). Deduced protein of 152 amino acids (16.

Biochemical and molecular characterization of stress-induced β-carbonic anhydrase from a C(4) plant, Pennisetum glaucum.

Genes encoding for many β-carbonic anhydrases and their functions in various developmental processes are well established in lower plants, however, similar studies are limited in higher plants. We report the cloning and characterization of cDNA encoding for a β-carbonic anhydrase (PgCA) from Pennisetum glaucum, a C(4) crop plant. cDNA encoding 249 amino acids and its deduced amino acid sequence analysis revealed that is related to other plant β-CA family members with an over all conserved architecture of a typical β-CA protein.

Molecular cloning and characterization of genes encoding Pennisetum glaucum ascorbate peroxidase and heat-shock factor: interlinking oxidative and heat-stress responses.

The recent genetic and biochemical studies reveal a considerable overlap among cellular processes in response to heat and oxidative stress stimuli in plants suggesting an intimate relationship between the heat-shock response and oxidative stress responses. Pennisetum glaucum (Pg) seedlings were exposed to heat stress (42 degrees C for 0.5, 1.

A high-throughput genome-walking method and its use for cloning unknown flanking sequences.

We developed a PCR-based high-throughput genome-walking protocol. The novelty of this protocol is in the random introduction of unique walker primer binding sites into different regions of the genome efficiently by taking advantage of the rolling circle mode of DNA synthesis by Phi29 DNA polymerase after annealing the partially degenerate primers to the denatured genomic DNA. The inherent strand-displacement activity of the Phi29 DNA polymerase displaces the 5' ends of downstream strands and DNA synthesis continues, resulting in a large number of overlapping fragments that cover the whole genome with the unique walker adapter attached to the 5' end of all the genomic DNA fragments.