title: Global analysis of gene expression in response to L-Cysteine deprivation in the anaerobic protozoan parasite Entamoeba histolytica.
abstract: BACKGROUND
  Entamoeba histolytica, an enteric protozoan parasite, causes amebic colitis and extra intestinal abscesses in millions of inhabitants of endemic areas. E. histolytica completely lacks glutathione metabolism but possesses L-cysteine as the principle low molecular weight thiol. L-Cysteine is essential for the structure, stability, and various protein functions, including catalysis, electron transfer, redox regulation, nitrogen fixation, and sensing for regulatory processes. Recently, we demonstrated that in E. histolytica, L-cysteine regulates various metabolic pathways including energy, amino acid, and phospholipid metabolism.
  
  
  RESULTS
  In this study, employing custom-made Affymetrix microarrays, we performed time course (3, 6, 12, 24, and 48 h) gene expression analysis upon L-cysteine deprivation. We identified that out of 9,327 genes represented on the array, 290 genes encoding proteins with functions in metabolism, signalling, DNA/RNA regulation, electron transport, stress response, membrane transport, vesicular trafficking/secretion, and cytoskeleton were differentially expressed (≥3 fold) at one or more time points upon L-cysteine deprivation. Approximately 60% of these modulated genes encoded proteins of no known function and annotated as hypothetical proteins. We also attempted further functional analysis of some of the most highly modulated genes by L-cysteine depletion.
  
  
  CONCLUSIONS
  To our surprise, L-cysteine depletion caused only limited changes in the expression of genes involved in sulfur-containing amino acid metabolism and oxidative stress defense. In contrast, we observed significant changes in the expression of several genes encoding iron sulfur flavoproteins, a major facilitator super-family transporter, regulator of nonsense transcripts, NADPH-dependent oxido-reductase, short chain dehydrogenase, acetyltransferases, and various other genes involved in diverse cellular functions. This study represents the first genome-wide analysis of transcriptional changes induced by L-cysteine deprivation in protozoan parasites, and in eukaryotic organisms where L-cysteine represents the major intracellular thiol.
authors.affiliation: Department of Parasitology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku, Tokyo 162-8640, Japan.
authors.forename: Afzal
authors.initials: A
authors.lastname: Husain
authors.forename: Ghulam
authors.initials: G
authors.lastname: Jeelani
authors.forename: Dan
authors.initials: D
authors.lastname: Sato
authors.forename: Tomoyoshi
authors.initials: T
authors.lastname: Nozaki
mesh_terms: D000693:Anaerobiosis; D046508:Culture Techniques; D003545:Cysteine / Q000378:metabolism*; D004748:Entamoeba histolytica / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism* / Q000469:parasitology; D005420:Flavoproteins / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D020868:Gene Silencing; D017125:Genes, Protozoan / Q000235:genetics; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D009696:Nucleic Acids / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D018384:Oxidative Stress / Q000235:genetics; D026361:Reactive Nitrogen Species / Q000235:genetics; D015398:Signal Transduction / Q000235:genetics
chemical_list: D005420:Flavoproteins; D026901:Membrane Transport Proteins; D009696:Nucleic Acids; D026361:Reactive Nitrogen Species; D003545:Cysteine