RESUMO
MicroRNAs (miRNAs) are small regulatory RNAs that play a significant role in eukaryotes by targeting mRNAs for cleavage or translational repression. Recent studies have also shown them to be associated with cellular changes following viral infection. Mink enteritis virus (MEV) is one of the most important viral pathogens in the mink industry. To study the involvement of miRNAs in the MEV infection process, we used Illumina's ultrahigh throughput approach to sequencing miRNA libraries from the feline kidney (F81) cell line before and after infection with MEV. Using this bioinformatics approach we identified 196 known mammalian miRNA orthologs belonging to 152 miRNA families in F81 cells. Additionally, 97 miRNA*s of these miRNAs were detected. As well as known miRNAs, 384 and 398 novel miRNA precursor candidates were identified in uninfected and MEV-infected F81 cells respectively that have not been reported in other mammals. In MEV-infected cells 3 miRNAs were significantly down-regulated and 4 up-regulated including 3 significantly. The majority (12 of 16) of randomly selected miRNA expression profiles by qRT-PCR were consistent with those identified by deep sequencing. A total of 88 miRNAs were predicted to target interferon-associated genes; 6 appear to target the 3'UTR of MEV-specific receptor transferring receptor mRNAs; and 8 to target the MEV mRNA coding region. No miRNAs coded by MEV itself were detected.
Assuntos
Biomarcadores/metabolismo , Panleucopenia Felina/genética , Perfilação da Expressão Gênica , Rim/metabolismo , MicroRNAs/genética , Enterite Viral do Vison/genética , Vírus da Enterite do Vison/patogenicidade , Animais , Gatos , Células Cultivadas , Biologia Computacional , Panleucopenia Felina/virologia , Rim/virologia , Enterite Viral do Vison/virologia , Análise de Sequência com Séries de Oligonucleotídeos , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Canine parvovirus type 2 (CPV) is a virulent pathogen that emerged in the late 1970s, probably originating from feline panleukopenia virus (FPLV) or a closely related carnivore parvovirus belonging to the feline parvovirus (FPV) subspecies. In contrast to FPLV, CPV has evolved rapidly since its emergence. The original antigenic type of CPV disappeared more than two decades ago and several new antigenic as well as genetic CPV variants have appeared and spread in the field. Both high mutation rate and positive selection of mutations in the capsid gene appear to be the driving force for such rapid evolution. In addition, genetic recombination has been assessed as a factor in parvovirus evolution. Recently, we provided the first evidence of inter-antigenic type recombination of CPV in nature. Here, an inter-FPV subspecies recombinant was revealed by analyzing the genetic data deposited in databases with several recombination detection programs, and by phylogeny. FPLV strain XJ-1, submitted by Su et al., Harbin, China in 2007 (GenBank accession no. EF988660), was most likely generated by recombination between CPV and FPLV. Its genome was generally composed of the NS1 gene of CPV origin and the VP1 gene of FPLV origin. This is the first demonstration of recombination between different FPV subspecies in nature. Consequently, recombination should be considered as an element in the generation and evolution of parvoviruses of the FPV subspecies.
