Han Xiao PhD thesis - Research@StAndrews:FullText - University of ...

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Fig. 1.5. Genetic map of a typical member of each genus of the Paramyxoviridae. The single-stranded, non-segmented, negative-sense RNA genome of paramyxoviruses consists of 6 to 10 tandem linked genes, depending on the species. The genes are shown as boxes that are drawn to approximate scale. These genes are separated by intergenic regions. Flanking the genome is a 3’ extracistronic region of ~50 nucleotides known as the leader and 5’ extracistronic region of 50 to 161 nucleotides known as the trailer.
Fig. 1.6. Schematic representation of the influenza A virus particle. Virions are decorated with two surface glycoproteins, HA and NA. M2 ion channel protein is also inserted into the lipid bilayer. Underneath the lipid envelope is M1 protein. The genome is composed of eight segments of single-stranded negativesense RNA genome encapsidated by the nucleoprotein. These ribonucleoproteins (RNPs) are also associated with components of polymerase complex (PB2, PB1, PA). NS1 and PB1-F2 are only found in infected cells, therefore they are not displayed in the diagram. The proteins encoded by each genomic segment are indicated. Diagram adapted from Lamb & Krug, 2001.
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VIRUS AND INTERFERON: A FIGHT FOR S
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Abstract The Interferon (IFN) famil
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3. Permission for electronic public
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Abbreviations ADAR1 Adenosine deami
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PI3K Phosphatidylinositol 3-kinase.
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1.2.2 Classification of paramyxovir
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2.5.1 RNA extraction……………
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1.1 The Interferon system The inter
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structural homologies: they both co
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1.1.2 Mechanisms of IFN signalling
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promoters, whilst others have both
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actively regulates diverse cellular
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2007)]. Mx1 gene expression is norm
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(Hsiang, Zhao, and Krug, 2009; Lai
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1.1.4 Virus encoded IFN antagonists
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1.1.5 Virus shut-off of host protei
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sector. The aetiological agent is N
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1.2.3 Virion structure The virions
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of P and V proteins are unique. 23
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species. The expression of P/V/C pr
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al., 1992; Riedl et al., 2002). M p
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forming the postfusion hairpin conf
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antigenically distinct HA subtypes
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asic amino acids sequence R-X-R/K-R
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egins with PB1 binding to the 5’
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stage of replication, full-length p
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into infectious virion. Finally, co
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with a number of host proteins invo
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infection, NS1 is found not only in
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1.3.4.3 NS1 and host immune respons
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The precise mechanism for how NS1 i
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CPSF30 and PABPII, some IFN-β pre-
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2008; Shin et al., 2007). Recently,
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2.1 Mammalian cells and tissue cult
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supplemented with Roferon recombina
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10. rUd-103/106B: The NS1 protein c
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Confluent monolayers of cells were
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2.3 DNA subcloning 2.3.1 cDNA synth
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appropriate time and temperature fo
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2.4.3 Immunofluorescence For immuno
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2.4.4 Preparation of radio-labelled
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emove unincorporated DIG nucleotide
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2.6 Miscellaneous assays 2.6.1 Lent
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Antibodies Antibodies were used in
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2.8 Primers PRIMERS DESCRIPTION SEQ
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2.9 Plasmids PLASMIDS pMD-VSVG pCMV
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A549 cells is that, IFN released fr
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3.1.3 Study of the mechanism of hos
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3.1.4 PIV5 can dismantle the IFN-in
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the majority of cells remained nega
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3.1.6 Influenza virus and the IFN-i
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these events occur only at the begi
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were detecting only the incoming vR
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naïve A549 cells and MxA-knockdown
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3.2.6 Effects of introducing MxA ba
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expression. However, over-expressio
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expression of MxA alone is insuffic
Page 115 and 116: numbers both around 100-fold, sugge
Page 117 and 118: 3.3.4 IFNγ does not have a signifi
Page 119 and 120: 4.2 Comparison of the mechanisms of
Page 121 and 122: 4.3 Influenza A virus shut-off of h
Page 123 and 124: 4.5 MxA recognition of NP protein E
Page 125 and 126: ability of IFN to block the nuclear
Page 127 and 128: Akira, S., Uematsu, S., and Takeuch
Page 129 and 130: Bonjardim, C. A. (2005). Interferon
Page 131 and 132: Cuddihy, A. R., Li, S., Tam, N. W.,
Page 133 and 134: Falvo, J. V., Thanos, D., and Mania
Page 135 and 136: Greenspan, D., Palese, P., and Krys
Page 137 and 138: Hirano, A., Wang, A. H., Gombart, A
Page 139 and 140: Jiang, D., Guo, H., Xu, C., Chang,
Page 141 and 142: Landis, H., Simon-Jodicke, A., Klot
Page 143 and 144: Martin, J., Wharton, S. A., Lin, Y.
Page 145 and 146: Neumann, G., Hughes, M. T., and Kaw
Page 147 and 148: (NAT) 2 acetylator status and age o
Page 149 and 150: Rodriguez, A., Perez-Gonzalez, A.,
Page 151 and 152: Staeheli, P., Pitossi, F., and Pavl
Page 153 and 154: Uze, G., and Monneron, D. (2007). I
Page 155 and 156: Wong, A. H., Tam, N. W., Yang, Y. L
Page 157 and 158: A.1 Plasmid map of the pLKO.1 with
Page 159 and 160: A.2 Plasmid map of the pdlNotI’IR
Page 161 and 162: Virus Mutation Comments rUd-NS1-R38
Page 163 and 164: Fig. 1.2. Schematic representation
Page 165: Fig. 1.4. Schematic representation
Page 169 and 170: (A) (B) Fig. 1.8. Replication and t
Page 171 and 172: (A) (B) Hours p.i. Mock PIV5 12 18
Page 173 and 174: Mock 103/106 rUd-184-8(L) rUd-184-8
Page 175 and 176: -antibody +antibody (A) (B) 16h (C)
Page 177 and 178: (A) 24h 48h -IFN +IFN (B) NS1 MxA M
Page 179 and 180: (A) 50 Percentage of cells positive
Page 181 and 182: (A) -IFN +IFN vRNA Merge vRNA Merge
Page 183 and 184: -IFN +IFN NP Merge NP Merge Mock A5
Page 185 and 186: Mock A549 A549-MxA -IFN +IFN Fig.3.
Page 187 and 188: (A) NP MxA Merge -IFN Mock +IFN (B)
Page 189 and 190: (A) A549 A549-MxA shMxA -7 -7 -7 -I
Page 191 and 192: (A) Hep2 Hep2/Npro Hep2/shISG56 (B)
Page 193: Fig. 3.22. IFNγ does not have an o
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