title: Transcriptional profiling of Entamoeba histolytica trophozoites.
abstract: We have developed an Entamoeba histolytica genomic DNA microarray and used it to develop a transcriptional profile of 1,971 E. histolytica (HM-1:IMSS) genes. The arrays accurately detected message abundance and 31-47% of amebic genes were expressed under standard tissue culture conditions (levels detectable by Northern blot analysis or RT-PCR respectively). Genes expressed at high levels ( approximately 2% of total) included actin (8.m00351), and ribosomal genes (20.m00312). Moderately expressed genes ( approximately 14% of total) included cysteine proteinase (191.m00117), profilin (156.m00098), and an Argonaute family member (11.m00378). Genes with low-level expression ( approximately 15% of total) included Ariel1 (160.m00087). Genes with very low expression ( approximately 16% of total) and those not expressed ( approximately 52% of total) included encystation-specific genes such as Jacob cyst wall glycoprotein (33.m00261), chitin synthase (3.m00544), and chitinase (22.m00311). Transcriptional modulation could be detected using the arrays with 17% of genes upregulated at least two-fold in response to heat shock. These included heat shock proteins (119.m00119 and 279.m00091), cyst wall glycoprotein Jacob (33.m00261), and ubiquitin-associated proteins (16.m00343; 195.m00092). Using Caco-2 cells to model the host-parasite interaction, we verified that host cell killing was dependent on live ameba. However, surprisingly these events did not appear to induce major transcriptional changes in the parasites.
authors.affiliation: Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, CA 94305-5124, USA.
authors.forename: Ryan C
authors.initials: RC
authors.lastname: MacFarlane
authors.forename: Preetam H
authors.initials: PH
authors.lastname: Shah
authors.forename: Upinder
authors.initials: U
authors.lastname: Singh
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D018938:Caco-2 Cells; D004748:Entamoeba histolytica / Q000235:genetics* / Q000502:physiology; D004749:Entamoebiasis / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D017125:Genes, Protozoan; D006790:Host-Parasite Interactions; D006358:Hot Temperature; D006801:Humans; D008018:Life Cycle Stages; D020411:Oligonucleotide Array Sequence Analysis; D015800:Protozoan Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: D015800:Protozoan Proteins