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      Effect of siRNA mediated suppression of signaling lymphocyte activation molecule on replication of peste des petits ruminants virus in vitro

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          Abstract

          Signaling lymphocyte activation molecule (SLAM) expression was inhibited in B95a cell line using siRNA and the effect of SLAM inhibition on peste des petits ruminants virus (PPRV) replication and infectivity titre was studied. SLAM suppression was assessed using real-time PCR and flow cytometry to confirm suppression at the m-RNA and protein levels, respectively. Three chemically synthesized siRNAs were transfected individually using oligofectamine into B95a cell line. This resulted in SLAM suppression from 48 to 454-folds, in comparison to the untransfected B95a cell line. When the SLAM suppressed B95a cell line was infected with PPRV, replication was reduced by 12–143-folds and virus titre was reduced from log 10 1.09 to 2.28. siRNA 3 showed the most potent inhibition of SLAM expression both at m-RNA and protein levels. This also caused the maximum reduction of virus replication and virus titre. A 100-fold reduction in PPRV titres was seen in anti-SLAM antibody neutralized B95a cell line. This further confirms that SLAM is one of the (co) receptors for PPRV. However, the presence of other putative virus receptor(s) is/are not ruled out.

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          Guidelines for the selection of highly effective siRNA sequences for mammalian and chick RNA interference.

          Kumiko Ui-Tei, Yuki Naito, Fumitaka Takahashi … (2004)
          In the present study, the relationship between short interfering RNA (siRNA) sequence and RNA interference (RNAi) effect was extensively analyzed using 62 targets of four exogenous and two endogenous genes and three mammalian and Drosophila cells. We present the rules that may govern siRNA sequence preference and in accordance with which highly effective siRNAs essential for systematic mammalian functional genomics can be readily designed. These rules indicate that siRNAs which simultaneously satisfy all four of the following sequence conditions are capable of inducing highly effective gene silencing in mammalian cells: (i) A/U at the 5' end of the antisense strand; (ii) G/C at the 5' end of the sense strand; (iii) at least five A/U residues in the 5' terminal one-third of the antisense strand; and (iv) the absence of any GC stretch of more than 9 nt in length. siRNAs opposite in features with respect to the first three conditions give rise to little or no gene silencing in mammalian cells. Essentially the same rules for siRNA sequence preference were found applicable to DNA-based RNAi in mammalian cells and in ovo RNAi using chick embryos. In contrast to mammalian and chick cells, little siRNA sequence preference could be detected in Drosophila in vivo RNAi.
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            Sialic acid species as a determinant of the host range of influenza A viruses.

            Barbara Holland, M Kiso, T. Ito … (2000)
            The distribution of sialic acid (SA) species varies among animal species, but the biological role of this variation is largely unknown. Influenza viruses differ in their ability to recognize SA-galactose (Gal) linkages, depending on the animal hosts from which they are isolated. For example, human viruses preferentially recognize SA linked to Gal by the alpha2,6(SAalpha2,6Gal) linkage, while equine viruses favor SAalpha2,3Gal. However, whether a difference in relative abundance of specific SA species (N-acetylneuraminic acid [NeuAc] and N-glycolylneuraminic acid [NeuGc]) among different animals affects the replicative potential of influenza viruses is uncertain. We therefore examined the requirement for the hemagglutinin (HA) for support of viral replication in horses, using viruses whose HAs differ in receptor specificity. A virus with an HA recognizing NeuAcalpha2,6Gal but not NeuAcalpha2,3Gal or NeuGcalpha2,3Gal failed to replicate in horses, while one with an HA recognizing the NeuGcalpha2,3Gal moiety replicated in horses. Furthermore, biochemical and immunohistochemical analyses and a lectin-binding assay demonstrated the abundance of the NeuGcalpha2,3Gal moiety in epithelial cells of horse trachea, indicating that recognition of this moiety is critical for viral replication in horses. Thus, these results provide evidence of a biological effect of different SA species in different animals.
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              Regulation of ATR substrate selection by Rad17-dependent loading of Rad9 complexes onto chromatin.

              Lee Zou, David Cortez, Stephen J. Elledge (2002)
              Cells respond to DNA damage by activating a network of signaling pathways that control cell cycle progression and DNA repair. Genetic studies in yeast suggested that several checkpoint proteins, including the RFC-related Rad17 protein, and the PCNA-related Rad1-Rad9-Hus1 protein complex might function as sensors of DNA damage. In this study, we show that the human Rad17 protein recruits the Rad9 protein complex onto chromatin after damage. Rad17 binds to chromatin prior to damage and is phosphorylated by ATR on chromatin after damage but Rad17's phosphorylation is not required for Rad9 loading onto chromatin. The chromatin associations of Rad17 and ATR are largely independent, which suggests that they localize to DNA damage independently. Furthermore, the phosphorylation of Rad17 requires Hus1, suggesting that the Rad1-Rad9-Hus1 complex recruited by Rad17 enables ATR to recognize its substrates. Our data are consistent with a model in which multiple checkpoint protein complexes localize to sites of DNA damage independently and interact to trigger the checkpoint-signaling cascade.
              Article 0 comments Cited 75 times – based on 0 reviews     Review now
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                Author and article information

                Contributors
                G. Dhinakar Raj
                Journal
                Journal ID (nlm-ta): Virus Res
                Journal ID (iso-abbrev): Virus Res
                Title: Virus Research
                Publisher: Elsevier B.V.
                ISSN (Print): 0168-1702
                ISSN (Electronic): 1872-7492
                Publication date PMC-release: 11 June 2008
                Publication date (Print): September 2008
                Publication date (Electronic): 11 June 2008
                Volume: 136
                Issue: 1
                Pages: 118-123
                Affiliations
                [a ]Department of Animal Biotechnology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600051, India
                [b ]Department of Veterinary Pathology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai 600051, India
                Author notes
                [* ]Corresponding author. Tel.: +91 44 25381506; fax: +91 44 25369301. dhinakarraj@ 123456yahoo.com
                Article
                Publisher ID: S0168-1702(08)00178-0
                DOI: 10.1016/j.virusres.2008.04.026
                PMC ID: 7127705
                PubMed ID: 18550191
                SO-VID: df658d06-ee04-4a20-96b5-4a289b1bd067
                Copyright statement: Copyright © 2008 Elsevier B.V. All rights reserved.
                License:

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                Date received : 1 February 2008
                Date revision received : 25 April 2008
                Date accepted : 27 April 2008
                Categories
                Subject: Article

                ScienceOpen disciplines: Microbiology & Virology
                Keywords: pprv, peste des petits ruminants virus,sirna, small interfering rna,slam, signaling lymphocyte activation molecule,peste des petits ruminants,sirna,transfection,signaling lymphocyte activation molecule,b95a cell line,real-time pcr,flow cytometry
                Data availability:
                ScienceOpen disciplines: Microbiology & Virology
                Keywords: pprv, peste des petits ruminants virus, sirna, small interfering rna, slam, signaling lymphocyte activation molecule, peste des petits ruminants, sirna, transfection, signaling lymphocyte activation molecule, b95a cell line, real-time pcr, flow cytometry

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