small rnas with 5′-polyphosphate termini associate with a piwi-related protein and regulate gene expression in the single-celled eukaryote entamoeba histolytica小rna 5u2032多磷酸盐末端与piwi-related蛋白和调节基因的表达在单细胞真核痢疾阿米巴.pdf

Small RNAs with 59-Polyphosphate Termini Associate with a Piwi-Related Protein and Regulate Gene Expression in the Single-Celled Eukaryote Entamoeba histolytica 1 1 2 1¤ 1,2 Hanbang Zhang , Gretchen M. Ehrenkaufer , Justine M. Pompey , Jason A. Hackney , Upinder Singh * 1 Division of Infectious Diseases, Department of Internal Medicine, Stanford University School of Medicine, Stanford, California, United States of America, 2 Department of Microbiology and Immunology, Stanford University School of Medicine, Stanford, California, United States of America Abstract Small interfering RNAs regulate gene expression in diverse biological processes, including heterochromatin formation and DNA elimination, developmental regulation, and cell differentiation. In the single-celled eukaryote Entamoeba histolytica, we have identified a population of small RNAs of 27 nt size that (i) have 59-polyphosphate termini, (ii) map antisense to genes, and (iii) associate with an E. histolytica Piwi-related protein. Whole genome microarray expression analysis revealed that essentially all genes to which antisense small RNAs map were not expressed under trophozoite conditions, the parasite stage from which the small RNAs were cloned. However, a number of these genes were expressed in other E. histolytica strains with an inverse correlation between small RNA and gene expression level, suggesting that these small RNAs mediate silencing of the cognate gene. Overall, our results demonstrate that E. histolytica has an abundant 27 nt small RNA population, with features similar to secondary siRNAs from C. elegans, and which appear to regulate gene expression. These data