RNA-Seq-Based Metatranscriptomic and Microscopic Investigation Reveals Novel Metalloproteases of Neobodo sp. as Potential Virulence Factors for Soft Tunic Syndrome in Halocynthia roretzi

摘要: Bodonids and trypanosomatids are derived from a common ancestor with the bodonids being more primitive lineage. The Neobodonida, one of three clades bodonids, can be free-living, commensal or parasitic. Despite ecological evolutionary significance these organisms, however, many their biological pathological features currently unknown. Here, we employed metatranscriptomics using RNA-seq technology combined field-emission microscopy to reveal virulence factors recently described genus Neobodonida that is considered responsible for ascidian soft tunic syndrome (AsSTS), but whose pathogenesis unclear. Our microscopic observation infected tissues suggested putative factors, enabling us extract novel candidate transcripts; included cysteine proteases families C1 C2, serine S51 S9 families, metalloproteases grouped into M1, M3, M8, M14, M16, M17, M24, M41, M49. Protease activity/inhibition assays estimation expression levels within gene clusters allowed identify metalloprotease-like enzymes as potential attributes AsSTS. Furthermore, multimarker-based phylogenetic analysis 1,184 concatenated amino acid sequences clarified order Neobodo sp. In sum, herein used elucidate in situ profiles uncharacterized transcripts sp., results select genes relevant pathogenesis, empirical screening define important factors.