Ribosomal Frameshifting of Trichomonas vaginalis virus 2, 3, and 4 Isolates

Trichomoniasis, the most common non‐viral sexually transmitted disease with an estimated 2 million new infections every year in the world, caused by a flagellated protozoan parasite Trichomonas vaginalis . T. vaginalis itself harbors up to four strains of non‐segmented double stranded RNA Trichomonas vaginalis virus (TVV1–4). T. vaginalis virus encodes two genes, an upstream capsid open reading frame (CP ORF) and a downstream fusion RNA‐dependent RNA polymerase (RdRp ORF). Literature studies suggest TVV2 and TVV3 might utilize −1 bp ribosomal frameshifting, while there is strong evidence that TVV1 undergoes −2 bp frameshifting during translation of RdRp in fusion with its capsid. However, there are not many available known genome sequences or many studies done on TVV4 ribosomal frameshifting. Here, we begin to investigate TVVs 2–4. Methods We used the only three known consensus sequence of TVV4, and designed sequencing primers. We also used cell cultivation, ultracentrifugation, mass spectrometry, RNA isolation, SDS‐PAGE analysis, RT PCR, qRT PCR, gel electrophoresis, and DNA isolation and purification. Results The genome of a clinical isolate infected with TVV4 has been nearly completed. Virions were isolated by ultracentrifugation. Conclusion New samples of TVV4 genome have been sequenced in this research for future studies, including LC‐MS analysis of tryptic digests that will allow elucidation of the frameshifting mechanism in TVVs 2–4. Support or Funding Information This work was supported by the Mississippi INBRE, funded by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under grant number P20GM103476