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“The influence of miRNAs on the host-pathogen environment is largely unknown and under intensive investigation. Whether produced by the pathogen or by the host cell, these miRNAs will sculpt the intracellular landscape, as their activity will ultimately affect levels of target proteins. Using a high-throughput sequencing approach, we identified 19 novel small RNAs produced during the early hours of herpes simplex virus type 1 (HSV-1) infection in epithelial cells. Six of the novel RNAs had predicted folds characteristic of miRNAs. One of the six, miR-92944, which resides in the 5′ UTR of the ul42 gene in the sense orientation, was confirmed buy Ro-3306 as a bona
fide miRNA by RT-PCR and stem-loop PCR analysis. Northern blot analysis was used to observe the precursor forms of miR-92944. Viral mutants that do not produce miR-92944 exhibited significant reductions in viral titers in both single and multi-step growth analysis and a fourfold reduction in plaque size. The miR-92944 mutants produce wild-type levels of ICP4, UL42, VP5, and gC proteins contain no additional changes in the DNA sequence surrounding the site of mutagenesis. The defective phenotype of miR-92944 mutants was complemented in V42.3 cells, which contain
the 5′UTR of ul42. We also found that miR-H1 expression was diminished in cells infected with the miR-92944 mutant virus. This study provides new information on the miRNA landscape during the early stages of HSV-1 infection and reveals novel U0126 clinical trial targets for antagonistic molecules that may curtail the establishment of lytic or latent virus infection.”
“Concatenated sequence alignments are often used to infer species-level relationships.
Previous studies have shown that analysis of concatenated data using maximum likelihood (ML) can produce misleading results when loci have differing gene tree topologies due to incomplete lineage sorting. Here, we develop a polynomial time method that utilizes the modified mincut supertree algorithm to construct an estimated species tree from inferred rooted triples of concatenated alignments. We term this method SuperMatrix Rooted Triple (SMRT) Selleckchem Sonidegib and use the notation SMRT-ML when rooted triples are inferred by ML. We use simulations to investigate the performance of SMRT-ML under Jukes-Cantor and general time-reversible substitution models for four- and five-taxon species trees and also apply the method to an empirical data set of yeast genes. We find that SMRT-ML converges to the correct species tree in many cases in which ML on the full concatenated data set fails to do so. SMRT-ML can be conservative in that its output tree is often partially unresolved for problematic clades.