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

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Fig. 1.3. Type I and type II IFN signalling pathways. Type I signalling is initiated by type I IFNs binding to a common receptor at the cell surface. This leads to the activation of the receptor-associated tyrosine kinases JAK1 and Tyk2, which phosphorylate STATs. Phosphorylated STAT1 and STAT2 form a heterodimer with each other by recognizing SH2 domains, and then translocated into the nucleus, where it interacts with the DNA-binding protein IRF-9, forming ISGF3. ISGF3 binds to a cis element, ISRE, located on in the promoter region of most ISGs. Type II IFN binds a distinct receptor, composed of IFNGR1 and IFNGR2, which are associated with JAK1 and JAK2, respectively. Activated JAKs phosphorylate STAT1, inducing the formation of STAT1 homodimers that translocate to the nucleus and bind to GAS elements of certain ISGs, thereby initiating their transcription (Diagram adapted from Platanias, 2005).
Fig. 1.4. Schematic representation of the Parainfluenza virus type 5 (PIV5) virion structure. The spherical particle of PIV5 consists of an outer lipid membrane with membrane protein hemagglutinin-neuraminidase (HN) attachment glycoprotein, and the fusion (F) glycoprotein. The small integral membrane protein, SH, is unique for PIV5. Underneath the lipid bilayer is matrix protein, essential for maintaining virus integrity. Inside the virion is the single-stranded, negative-sense RNA genome encapsidated by nucleocapsids protein (NP). Associated with NP are the L and P proteins, which have the RNA-dependent RNA polymerase activity. V protein of PIV5 is found as a virion component, whilst V proteins of other members of paramyxoviruses are found only in virus-infected cells.
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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
Page 113 and 114: 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: Fig. 1.2. Schematic representation
Page 167 and 168: Fig. 1.6. 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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