Poliovirus Protein 3A Binds and Deregulates LIS1, Causing Block of Membrane Protein Trafficking and Deregulation of Cell Division

Abstract

Many viruses encode anti-apoptotic proteins that have been used as valuable tools for identification and analysis of key cellular regulators of programmed cell death. Here we demonstrate that the poliovirus protein 3A, previously shown to exhibit anti-apoptotic activity, binds and inactivates LIS1, a component of the dynein/dynactin motor complex, encoded by the gene mutated in patients with type I lissencephaly ("smooth brain"), thereby causing deregulation of endoplasmatic reticilum-to-Golgi vesicular transport, resulting in rapid disappearance of short-living receptors from the plasma membrane and loss of cell sensitivity to TNF and interferon. Truncated derivatives of LIS1, acting in a dominant negative manner, cause similar effects. However, 3A, being an endoplasmic reticulumbound protein, locks Golgi-targeted YFP in the endoplasmatic reticilum, while expression of LIS1 mutants results in a dispersed cytoplasmic localization of the reporter protein. LIS1 dysfunction caused by ectopic expressing 3A or LIS1 mutants, as well as by overexpression of wild type LIS1, leads to cell blocking at the postmitotic stage associated with inability to undergo cytokinesis. Thus, the use of poliovirus protein as a research tool allowed us to reveal the role of cellular protein LIS1 in membrane protein trafficking, maintenance of Golgi integrity, surface presentation of unstable receptors, cell sensitivity to TNF-induced apoptosis and cell cycle progression.

Functional interplay between LIS1, NDE1 and NDEL1 in dynein-dependent organelle positioning

“…However, the requirement for LIS1 in these functions is controversial, despite its involvement in other dynein-dependent cellular processes. We found that simple overexpression of GFP-LIS1 in HeLaM cells did not cause Golgi complex dispersal (a typical phenotype seen following disruption of dynein activity), in agreement with some studies (Dujardin et al, 2003;Faulkner et al, 2000;Tai et al, 2002), but in contrast to other work (Kondratova et al, 2005;Sasaki et al, 2000). Instead, the Golgi became somewhat more compact compared with control cells (Fig.…”

“…Drosophila LIS1 -/-neurons display defects in axonal transport, although the phenotype differs somewhat from DHC mutants (Liu et al, 2000). Overexpression of LIS1 has been reported to leave the Golgi complex position unchanged (Faulkner et al, 2000), more compact or dispersed (Kondratova et al, 2005). There is stronger evidence for an involvement of NDEL1 in membrane positioning, because overexpression of LIS1-or dynein-binding NDEL1 mutants alters the position and centripetal movement of a range of organelles and membrane cargo (Liang et al, 2004).…”

“…In several studies, interventions that inhibited other LIS1-dependent activities did not affect membrane organisation (Dujardin et al, 2003;Faulkner et al, 2000;Tai et al, 2002). Elsewhere, mild (Sasaki et al, 2000;Smith et al, 2000) or more severe (Ding et al, 2009;Kondratova et al, 2005;Rehberg et al, 2005) effects on Golgi positioning were seen upon LIS1 reduction, Summary LIS1, NDE1 and NDEL1 modulate cytoplasmic dynein function in several cellular contexts. However, evidence that they regulate dynein-dependent organelle positioning is limited.…”

Poliovirus infection blocks ERGIC-to-Golgi trafficking and induces microtubule-dependent disruption of the Golgi complex

“…Viral protein 3A has been shown to block ER-to-Golgi traffic on its own (Doedens and Kirkegaard, 1995) at the ERGIC-to-Golgi step (Wessels et al, 2005). This activity correlated with direct interactions with cellular proteins Lis1 (Kondratova et al, 2005) and GBF1 (Wessels et al, 2006). Notably, overexpression of cellular protein Arf1 or GBF1 abrogates the inhibition by coxsackie virus 3A of ER-to-Golgi traffic (Wessels et al, 2006).…”

Role of non-motile microtubule-associated proteins in virus trafficking

“…Moreover, some studies remain inconclusive as to whether virus-MAP interactions that stabilize MTs lead to enhanced infection by specifically affecting viral trafficking, or rather by affecting general transport. For instance, the poliovirus 3A protein was shown to bind to Lis1 and disrupt protein trafficking, but this was found to impact the cell surface expression of short-lived receptors, including those for tumor necrosis factor and interferon (IFN), rather than viral particle trafficking per se (102). Moreover, it has been reported that HIV-1 Tat also binds to Lis1 and this interaction might contribute to the effect of Tat on MT formation (103).…”