Kettering University

About: Kettering University is a education organization based out in Flint, Michigan, United States. It is known for research contribution in the topics: RNA & Antigen. The organization has 6842 authors who have published 7689 publications receiving 337503 citations. The organization is also known as: GMI Engineering & Management Institute & General Motors Institute.

Virus discovery by deep sequencing and assembly of virus-derived small silencing RNAs

Abstract: In response to infection, invertebrates process replicating viral RNA genomes into siRNAs of discrete sizes to guide virus clearance by RNA interference. Here, we show that viral siRNAs sequenced from fruit fly, mosquito, and nematode cells were all overlapping in sequence, suggesting a possibility of using siRNAs for viral genome assembly and virus discovery. To test this idea, we examined contigs assembled from published small RNA libraries and discovered five previously undescribed viruses from cultured Drosophila cells and adult mosquitoes, including three with a positive-strand RNA genome and two with a dsRNA genome. Notably, four of the identified viruses exhibited only low sequence similarities to known viruses, such that none could be assigned into an existing virus genus. We also report detection of virus-derived PIWI-interacting RNAs (piRNAs) in Drosophila melanogaster that have not been previously described in any other host species and demonstrate viral genome assembly from viral piRNAs in the absence of viral siRNAs. Thus, this study provides a powerful culture-independent approach for virus discovery in invertebrates by assembling viral genomes directly from host immune response products without prior virus enrichment or amplification. We propose that invertebrate viruses discovered by this approach may include previously undescribed human and vertebrate viral pathogens that are transmitted by arthropod vectors.

Hydroxyurea: inhibitory effect on dna metabolism.

Abstract: Hydroxyurea, hydroxyurethane, and dihydroxyurea inhibit incorporation of thymidine into the DNA of monolayers of HeLa cells. They do not affect incorporation of uridine into RNA or of leucine into protein. In contrast, hydroxylamine inhibits cellular incorporation of all three precursors: thymidine, uridine, and leucine. Hydroxyurea does not affect thymidine kinase, thymidylate kinase, or DNA polymerase reactions, but it does inhibit incorporation of cytidylic and guanylic acids into DNA in cell-free supernatants.