RESUMO
Rio Negro virophage (RNV) was co-isolated with a strain of mimivirus named sambavirus, from Brazilian Amazon. We report the near complete genome sequence of RNV, the first virophage isolated in Brazil. We also present new microscopical data demonstrating that RNV particles have similar dimensions to that described to sputnik virophages.
Assuntos
Acanthamoeba/virologia , Genoma Viral , Togaviridae/genética , Virófagos/genética , Brasil , Microscopia Eletrônica de Transmissão , Fases de Leitura Aberta , Filogenia , Togaviridae/isolamento & purificação , Togaviridae/ultraestrutura , Virófagos/isolamento & purificação , Virófagos/ultraestruturaRESUMO
The giant viruses are the largest and most complex viruses in the virosphere. In the last decade, new members have constantly been added to this group. Here, we provide an in-depth descriptive analysis of the replication cycle of Cedratvirus getuliensis, one of the largest viruses known to date. We tracked the virion entry, the early steps of virus factory and particles morphogenesis, and during this phase, we observed a complex and unique sequential organization of immature particle elements, including horseshoe and rectangular compartments, revealed by transverse and longitudinal sections, respectively, until the formation of the final ovoid-shaped striped virion. The genome and virion proteins are incorporated through a longitudinal opening in the immature virion, followed by the incorporation of the second cork and thickening of the capsid well. Moreover, many cell modifications occur during viral infection, including intense membrane trafficking important to viral morphogenesis and release, as evidenced by treatment using brefeldin A. Finally, we observed that Cedratvirus getuliensis particles are released after cellular lysis, although we obtained microscopic evidence that some particles are released by exocytosis. The present study provides new information on the unexplored steps in the life cycle of cedratviruses.
Assuntos
Vírus de DNA/fisiologia , Replicação Viral , Acanthamoeba castellanii/virologia , Citocalasinas/farmacologia , Citoplasma/efeitos dos fármacos , Citoplasma/virologia , Vírus de DNA/efeitos dos fármacos , Vírus de DNA/isolamento & purificação , Vírus de DNA/ultraestrutura , Exocitose , Estágios do Ciclo de Vida , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Esgotos/virologia , Vírion/ultraestrutura , Internalização do VírusRESUMO
For many years, gene expression in the three cellular domains has been studied in an attempt to discover sequences associated with the regulation of the transcription process. Some specific transcriptional features were described in viruses, although few studies have been devoted to understanding the evolutionary aspects related to the spread of promoter motifs through related viral families. The discovery of giant viruses and the proposition of the new viral order Megavirales that comprise a monophyletic group, named nucleo-cytoplasmic large DNA viruses (NCLDV), raised new questions in the field. Some putative promoter sequences have already been described for some NCLDV members, bringing new insights into the evolutionary history of these complex microorganisms. In this review, we summarize the main aspects of the transcription regulation process in the three domains of life, followed by a systematic description of what is currently known about promoter regions in several NCLDVs. We also discuss how the analysis of the promoter sequences could bring new ideas about the giant viruses' evolution. Finally, considering a possible common ancestor for the NCLDV group, we discussed possible promoters' evolutionary scenarios and propose the term "MEGA-box" to designate an ancestor promoter motif ('TATATAAAATTGA') that could be evolved gradually by nucleotides' gain and loss and point mutations.
Assuntos
Vírus Gigantes/genética , Regiões Promotoras Genéticas , Transcrição GênicaRESUMO
Acanthamoeba are natural hosts for giant viruses and their life cycle comprises two stages: a trophozoite and a cryptobiotic cyst. Encystment involves a massive turnover of cellular components under molecular regulation. Giant viruses are able to infect only the trophozoite, while cysts are resistant to infection. Otherwise, upon infection, mimiviruses are able to prevent encystment. This review highlights the important points of Acanthamoeba and giant virus interactions regarding the encystment process. The existence of an acanthamoebal non-permissive cell for Acanthamoeba polyphaga mimivirus, the prototype member of the Mimivirus genus, is analyzed at the molecular and ecological levels, and compared to a similar phenomenon previously described for Emiliana huxleyi and its associated phycodnaviruses: the 'Cheshire Cat' escape strategy.
Assuntos
Acanthamoeba/virologia , Vírus Gigantes/genética , Interações Hospedeiro-Patógeno/genética , Mimiviridae/genética , Encistamento de Parasitas/genética , Transdução de Sinais/genética , Trofozoítos/virologiaRESUMO
2'5'OAS are template-independent RNA polymerases with antiviral activity and important to homeostasis maintenance. Here we have developed quantitative PCR (qPCR) reactions for the detection of each individual 2'5'OAS human gene and used them to evaluate these gene levels in systemic sclerosis patients cells. The method was efficient for quantification of 2'5'OAS genes on human cells after interferon (IFN) treatment, and revealed that primary cells from patients with systemic sclerosis have increased basal levels of OASL and OAS2 genes. When treated, patients cells are able to induce all four 2'5'OAS genes. Our hypothesis is that abnormally circulating type I IFNs on the disease could be establishing a desensitized state on patients cells, making them refractory to subsequent IFN doses, and that OASL and OAS2 genes upregulation may be due to an IFN-independent stimulus. Further characterizing the biological activities of these genes, their induction pathways and their regulatory functions can lead to better understanding of systemic sclerosis molecular mechanisms and of their biological activities.
Assuntos
2',5'-Oligoadenilato Sintetase/genética , Leucócitos Mononucleares/imunologia , Reação em Cadeia da Polimerase/métodos , Escleroderma Sistêmico/genética , 2',5'-Oligoadenilato Sintetase/imunologia , Adulto , Idoso , Linhagem Celular Tumoral , Feminino , Expressão Gênica , Humanos , Interferon-alfa/farmacologia , Interferon beta/farmacologia , Isoenzimas/genética , Isoenzimas/imunologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/patologia , Masculino , Pessoa de Meia-Idade , Especificidade de Órgãos , Cultura Primária de Células , Escleroderma Sistêmico/imunologia , Escleroderma Sistêmico/patologia , Regulação para CimaRESUMO
ABSTRACT Rio Negro virophage (RNV) was co-isolated with a strain of mimivirus named sambavirus, from Brazilian Amazon. We report the near complete genome sequence of RNV, the first virophage isolated in Brazil. We also present new microscopical data demonstrating that RNV particles have similar dimensions to that described to sputnik virophages.
RESUMO
ABSTRACT Rio Negro virophage (RNV) was co-isolated with a strain of mimivirus named sambavirus, from Brazilian Amazon. We report the near complete genome sequence of RNV, the first virophage isolated in Brazil. We also present new microscopical data demonstrating that RNV particles have similar dimensions to that described to sputnik virophages.