dc.contributor.author | Guerrero Alonso, Ana | |
dc.date.accessioned | 2016-06-13T11:24:12Z | |
dc.date.available | 2016-06-13T11:24:12Z | |
dc.date.issued | 25/11/2016 | |
dc.date.submitted | 2016-06-13T12:10:16.151Z | |
dc.identifier.citation | Guerrero Alonso, A, 2016, Investigating early events in HIV-1 replication: the role of envelope signalling and PAF1 restriction factor, Queen Mary University of London. | en_US |
dc.identifier.uri | http://qmro.qmul.ac.uk/xmlui/handle/123456789/12823 | |
dc.description | MD (res) | en_US |
dc.description.abstract | To enter cells, HIV utilises the envelope (Env) protein to engage the CD4 receptor and a co-receptor, CXCR4 or CCR5. In addition to its role in HIV entry, the Env protein also triggers intracellular signalling events. Expression of the HIV-1 and HIV-2 Env gp41 cytoplasmic tails triggered a significant decrease in microtubule acetylation at the single cell level. Since microtubule stability is controlled by Rac1, the role of Rac1 in HIV-1 infection was investigated. Treatment of TZM-bl cells with a Rac1 inhibitor that blocks its upstream activators, Tiam1 and TrioN, inhibited HIV-1 NL4.3 (X4-tropic) but not HIV-1 BaL (R5-tropic) suggesting a role for these activators which is possibly, co-receptor dependent. Subsequently, an siRNA screen including 145 upstream regulators of Rac1, cdc42 and RhoA were investigated in their role in HIV-1 NL4.3 and HIV-1 BaL infection. As a result, 39 upstream regulators of Rho-GTPases were identified of which 5 proteins had effects on both HIV-1 BaL and HIV-1 NL4.3 infection which suggests a novel role of Rho-GTPase upstream regulators in HIV-1 infection. Early in infection, HIV-1 also encounters restriction factors such as RNA polymerase II-Associated Factor 1 (PAF1). The possible cytoplasmic or nuclear role of PAF1 in HIV-1 infection was investigated. Confocal imaging and cellular fractionation suggested that endogenous PAF1 is localised mostly in the nuclei but also in the cytoplasm. Overexpression of a murine PAF1 cytoplasm-localised mutant decreased HIV-1 infection similarly to wild-type overexpression. On the other hand, overexpression of a nuclear PAF1 construct decreased infection better than the wild-type. This suggests that PAF1 restricts HIV-1 via a nuclear function. In addition, infection time-course experiments revealed that HIV-1 counteracts the effect of PAF1 within 30 minutes of infection. Overall, the evidence presented, suggests that PAF1 has a nuclear mechanism and that its downregulation could take place in the cytoplasm. | en_US |
dc.language.iso | en | en_US |
dc.publisher | Queen Mary University of London | |
dc.subject | Medicine | en_US |
dc.subject | microRNAs | en_US |
dc.subject | smooth muscle cell differentiation | en_US |
dc.title | Investigating early events in HIV-1 replication: the role of envelope signalling and PAF1 restriction factor | en_US |
dc.type | Thesis | en_US |
dc.rights.holder | The copyright of this thesis rests with the author and no quotation from it or information derived from it may be published without the prior written consent of the author | |