title: Differential gene expression in Giardia lamblia under oxidative stress: significance in eukaryotic evolution.
abstract: Giardia lamblia is a unicellular, early branching eukaryote causing giardiasis, one of the most common human enteric diseases. Giardia, a microaerophilic protozoan parasite has to build up mechanisms to protect themselves against oxidative stress within the human gut (oxygen concentration 60 μM) to establish its pathogenesis. G. lamblia is devoid of the conventional mechanisms of the oxidative stress management system, including superoxide dismutase, catalase, peroxidase, and glutathione cycling, which are present in most eukaryotes. NADH oxidase is a major component of the electron transport chain of G. lamblia, which in concurrence with disulfide reductase, protects oxygen-labile proteins such as pyruvate: ferredoxin oxidoreductase against oxidative stress by sustaining a reduced intracellular environment. It also contains the arginine dihydrolase pathway, which occurs in a number of anaerobic prokaryotes, includes substrate level phosphorylation and adequately active to make a major contribution to ATP production. To study differential gene expression under three types of oxidative stress, a Giardia genomic DNA array was constructed and hybridized with labeled cDNA of cells with or without stress. The transcriptomic data has been analyzed and further validated using real time PCR. We identified that out of 9216 genes represented on the array, more than 200 genes encoded proteins with functions in metabolism, oxidative stress management, signaling, reproduction and cell division, programmed cell death and cytoskeleton. We recognized genes modulated by at least ≥ 2 fold at a significant time point in response to oxidative stress. The study has highlighted the genes that are differentially expressed during the three experimental conditions which regulate the stress management pathway differently to achieve redox homeostasis. Identification of some unique genes in oxidative stress regulation may help in new drug designing for this common enteric parasite prone to drug resistance. Additionally, these data suggest the major role of this early divergent ancient eukaryote in anaerobic to aerobic organism evolution.
authors: Division of Parasitology, National Institute of Cholera and Enteric Diseases, P-33, CIT Road, Scheme XM, Beliaghata, Kolkata 700010, India.
authors: Dibyendu
authors: D
authors: Raj
authors: Division of Parasitology, National Institute of Cholera and Enteric Diseases, P-33, CIT Road, Scheme XM, Beliaghata, Kolkata 700010, India.
authors: Esha
authors: E
authors: Ghosh
authors: Division of Parasitology, National Institute of Cholera and Enteric Diseases, P-33, CIT Road, Scheme XM, Beliaghata, Kolkata 700010, India.
authors: Avik K
authors: AK
authors: Mukherjee
authors: Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.
authors: Tomoyoshi
authors: T
authors: Nozaki
authors: Division of Parasitology, National Institute of Cholera and Enteric Diseases, P-33, CIT Road, Scheme XM, Beliaghata, Kolkata 700010, India. Electronic address: sandipanganguly@gmail.com.
authors: Sandipan
authors: S
authors: Ganguly
mesh_terms: D005075:Biological Evolution; D002455:Cell Division / Q000235:genetics; D004734:Energy Metabolism / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D016829:Giardia lamblia / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008954:Models, Biological; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000235:genetics*; D010101:Oxygen Consumption; D015203:Reproducibility of Results; D012098:Reproduction / Q000235:genetics
keywords: (reduced) nicotinamide adenine dinucleotide; (reduced) nicotinamide adenine dinucleotide phosphate; 2′,7′-dichlorodihydrofluorescein diacetate; 2′,7′-dichlorofluorescein; 4-(2-hydroxyethyl) piperazine-1-ethanesulfonic acid; ATP; DCF; GSH; Giardia; H(2)DCFDA; HEPES; NADH; NADPH; Oxidative stress; PBS; PCD; PCR; PCV; ROS; RT-PCR; SOD; SRP-64; SSC; TYIS-33; Transcriptomic; VSP; adenosine tri-phosphate; dTTP; dUTP; deoxythymidine triphosphate; deoxyuridine triphosphate; glutathione; phosphate buffered saline; polymerase chain reaction; programmed cell death protein like protein; protein for cell viability; reactive oxygen species; real time PCR; signal recognition particle-64; standard saline citrate; superoxide dismutase; tryptone–yeast extract–iron–serum-33; variant specific surface protein