Characterization of active ribosomal RNA harboring MITEs insertion in microsporidian Nosema bombycis genome.

Microsporidia are a group of obligate intracellular parasites of medical and agricultural importance, which can infect almost all animals, including human beings. Using the genome data of Nosema bombycis, four families of miniature inverted-repeat transposable elements (MITEs) in ribosomal DNA (rDNA) were characterized in the microsporidian N. bombycis and were named LSUME1, ITSME1, SSUME1, and SSUME2, respectively. The genome-wide investigation of these MITEs shows that these MITEs families distribute randomly in N. bombycis genome. All insertion sequences have conserved characteristics of MITEs, the direct repeat sequence and terminal inverted-repeat sequence at both ends of each MITEs sequence. Additionally, using the CLC RNA Workbench Software, secondary structures of rRNA containing MITEs sequence have been predicted and were located in variable region or expansion segment. Furthermore, using two different probes, one is prepared by MITE sequence only (short probe) and the other is prepared by MITE sequence flanking partial rDNA sequence (long probe); northern blotting and dot blotting have been performed to detect the transcriptional and functional activity of the rDNA containing MITEs insertion. Fortunately, we found that the rDNA, which harbors the MITE, not only can be transcripted but also can form a complete ribosome. This is an interesting thing that one gene can keep active even when it has been inserted with another sequence. But the biological and structural significance of this observation is not readily apparent.