PKA-dependent dynein switching from lysosomes to adenovirus: A novel form of host–virus competition

Scherer, Julian; Yi, Julie; Vallee, Richard B.

doi:10.1083/jcb.201307116

Cited by 64 publications

(89 citation statements)

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“…Internalized viral particles disrupt endosomal membranes, escape into the cytosol, and are transported along microtubules to the nuclear membrane, where viral DNA is imported into the cell nucleus followed by expression of its genetic program (66). Dynein-dynactin motors are switched away from Rab7-containing late endosomes and lysosomes to viral capsids in order to initiate transport to the nucleus by a mechanism involving a direct interaction between a hexon subunit of the capsid and dynein, leading to a partial inhibition of lysosome function (68). Consistent with these findings, we showed that Ad2 infection led to a significant reduction in cholesterol transport from late endosomes and lysosomes to other organelles by a mechanism that is normally masked by RID␣ expression in cells infected with wild-type Ad2.…”

Section: Discussionmentioning

confidence: 99%

Adenovirus Modulates Toll-Like Receptor 4 Signaling by Reprogramming ORP1L-VAP Protein Contacts for Cholesterol Transport from Endosomes to the Endoplasmic Reticulum

Cianciola

Chung

Manor

et al. 2017

J Virol

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Human adenoviruses (Ads) generally cause mild self-limiting infections but can lead to serious disease and even be fatal in high-risk individuals, underscoring the importance of understanding how the virus counteracts host defense mechanisms. This study had two goals. First, we wished to determine the molecular basis of cholesterol homeostatic responses induced by the early region 3 membrane protein RID␣ via its direct interaction with the sterol-binding protein ORP1L, a member of the evolutionarily conserved family of oxysterol-binding protein (OSBP)-related proteins (ORPs). Second, we wished to determine how this interaction regulates innate immunity to adenovirus. ORP1L is known to form highly dynamic contacts with endoplasmic reticulum-resident VAP proteins that regulate late endosome function under regulation of Rab7-GTP. Our studies have demonstrated that ORP1L-VAP complexes also support transport of LDL-derived cholesterol from endosomes to the endoplasmic reticulum, where it was converted to cholesteryl esters stored in lipid droplets when ORP1L was bound to RID␣. The virally induced mechanism counteracted defects in the predominant cholesterol transport pathway regulated by the late endosomal membrane protein Niemann-Pick disease type C protein 1 (NPC1) arising during early stages of viral infection. However, unlike NPC1, RID␣ did not reconstitute transport to endoplasmic reticulum pools that regulate SREBP transcription factors. RID␣-induced lipid trafficking also attenuated proinflammatory signaling by Toll-like receptor 4, which has a central role in Ad pathogenesis and is known to be tightly regulated by cholesterol-rich "lipid rafts." Collectively, these data show that RID␣ utilizes ORP1L in a way that is distinct from its normal function in uninfected cells to fine-tune lipid raft cholesterol that regulates innate immunity to adenovirus in endosomes.IMPORTANCE Early region 3 proteins encoded by human adenoviruses that attenuate immune-mediated pathology have been a particularly rich source of information regarding intracellular protein trafficking. Our studies with the early region 3-encoded RID␣ protein also provided fundamental new information regarding mechanisms of nonvesicular lipid transport and the flow of molecular information at membrane contacts between different organelles. We describe a new pathway that delivers cholesterol from endosomes to the endoplasmic reticulum, where it is esterified and stored in lipid droplets. Although lipid droplets are attracting renewed interest from the standpoint of normal physiology and human diseases, including those resulting from viral infections, experimental model systems for evaluating how and why they accumulate are still limited. Our studies also revealed an intriguing relationship between lipid droplets and innate immunity that may represent a new paradigm for viruses utilizing these organelles.KEYWORDS adenoviruses, cholesterol, endocytic pathway, endoplasmic reticulum, lipid droplet

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Section: Discussionmentioning

confidence: 99%

Adenovirus Modulates Toll-Like Receptor 4 Signaling by Reprogramming ORP1L-VAP Protein Contacts for Cholesterol Transport from Endosomes to the Endoplasmic Reticulum

Cianciola

Chung

Manor

et al. 2017

J Virol

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“…Similar effects of pathogens on trafficking adaptor proteins have been reported as well. Salmonella displaces RILP from Rab7-containing vesicles, enhancing kinesin-dependent trafficking (8), and adenovirus induces the phosphorylation of dynein to block the RILP-dynein binding (20). In each of these cases, the Rab protein and its adaptors are targets responsible for trafficking alterations.…”

Section: Discussionmentioning

confidence: 99%

Hepatitis C virus promotes virion secretion through cleavage of the Rab7 adaptor protein RILP

Wozniak

Long

Jones-Jamtgaard

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Hepatitis C virus (HCV) is an enveloped RNA virus that modifies intracellular trafficking processes. The mechanisms that HCV and other viruses use to modify these events are poorly understood. In this study, we observed that two different RNA viruses, HCV and Sendai, cause inhibition of ras-related protein Rab-7 (Rab7)-dependent endosome-lysosome fusion. In both cases, viral infection causes cleavage of the Rab7 adaptor protein RILP (Rab interacting lysosomal protein), which is responsible for linking Rab7 vesicles to dynein motor complexes. RILP cleavage results in the generation of a cleaved RILP fragment (cRILP) missing the N terminus of the molecule. Although RILP localizes in a perinuclear fashion, cRILP moves to the cell periphery. Both knockdown of RILP and expression of cRILP reproduced the HCV-induced trafficking defect, and restoring full-length RILP reversed the trafficking effects of virus. For the first 3 d after electroporation of HCV RNA, intracellular virus predominates over secreted virus, but the quantity of intracellular virus then rapidly declines as secreted virus dominates. The transition from the intracellular-predominant to the secretion-predominant phenotype corresponds to the time course of cRILP generation. Expressing cRILP directly prevents intracellular virus accumulation at early times without affecting net virus production. The ability of cRILP to promote virus secretion could be prevented by a kinesin inhibitor. HCV thus modifies cellular trafficking by cleaving RILP, which serves to redirect Rab7-containing vesicles to a kinesindependent trafficking mode promoting virion secretion. Cleavage of a Rab adaptor protein is thus a mechanism by which viruses modify trafficking patterns of infected cells.T he hepatitis C virus (HCV) is a major cause of chronic hepatitis, cirrhosis, and hepatocellular carcinoma. HCV is a positive-stranded RNA virus. It replicates on modified ER membranes (1), and virions are subsequently assembled on the surface of ER-associated lipid droplets (2). The processes necessary for virion trafficking to the plasma membrane and exocytosis have not been well defined. There is considerable evidence that HCV alters several membrane trafficking steps. It rearranges the ER to form the membranous web replication complex (3, 4), and it suppresses the fusion of autophagosomes and lysosomes, resulting in the accumulation of autophagic vacuoles (5). Although the mechanisms responsible for HCVinduced membrane rearrangements are poorly understood, it is logical to assume that redirection of vesicle trafficking pathways by HCV serve the viral lifecycle by initiating genome replication, preventing virion degradation, and optimizing virion secretion.A key molecule in the regulation of membrane trafficking is rasrelated protein Rab-7 (Rab7), a member of the small GTPase family that acts as a switch triggering the assembly of vesicle microtubule motor protein linkage complexes (6). Rab7 facilitates the maturation of endosomes and autophagosomes and also promotes the microtubule-...

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“…We relied on automated particle detection (25-nm resolution), tracking, and motility analysis software to determine segments of constant capsid velocities (16,24) (Fig. 4; see also Materials and Methods).…”

Section: Fig 2 Pk15-mng-vp26 Cells Produce Dual-color Prv180g (A) Timentioning

confidence: 99%

Dual-Color Herpesvirus Capsids Discriminate Inoculum from Progeny and Reveal Axonal Transport Dynamics

Scherer

Yaffe

Vershinin

et al. 2016

J Virol

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Alphaherpesviruses such as herpes simplex virus and pseudorabies virus (PRV) are neuroinvasive double-stranded DNA (dsDNA) viruses that establish lifelong latency in peripheral nervous system (PNS) neurons of their native hosts. Following reactivation, infection can spread back to the initial mucosal site of infection or, in rare cases, to the central nervous system, with usually serious outcomes. During entry and egress, viral capsids depend on microtubule-based molecular motors for efficient and fast transport. In axons of PNS neurons, cytoplasmic dynein provides force for retrograde movements toward the soma, and kinesins move cargo in the opposite, anterograde direction. The dynamic properties of virus particles in cells can be imaged by fluorescent protein fusions to the small capsid protein VP26, which are incorporated into capsids. However, single-color fluorescent protein tags fail to distinguish the virus inoculum from progeny. Therefore, we established a dual-color system by growing a recombinant PRV expressing a red fluorescent VP26 fusion (PRV180) on a stable cell line expressing a green VP26 fusion (PK15-mNG-VP26). The resulting dual-color virus preparation (PRV180G) contains capsids tagged with both red and green fluorescent proteins, and 97% of particles contain detectable levels of mNeonGreen (mNG)-tagged VP26. After replication in neuronal cells, all PRV180G progeny exclusively contain monomeric red fluorescent protein (mRFP)-VP26-tagged capsids. We used PRV180G for an analysis of axonal capsid transport dynamics in PNS neurons. Fast dual-color total internal reflection fluorescence (TIRF) microscopy, single-particle tracking, and motility analyses reveal robust, bidirectional capsid motility mediated by cytoplasmic dynein and kinesin during entry, whereas egressing progeny particles are transported exclusively by kinesins. IMPORTANCE Alphaherpesviruses are neuroinvasive viruses that infect the peripheral nervous system (PNS) of infected hosts as an integral part of their life cycle. Establishment of a quiescent or latent infection in PNS neurons is a hallmark of most alphaherpesviruses.Spread of infection to the central nervous system is surprisingly rare in natural hosts but can be fatal. Pseudorabies virus (PRV) is a broad-host-range swine alphaherpesvirus that enters neuronal cells and utilizes intracellular transport processes to establish infection and to spread between cells. By using a virus preparation with fluorescent viral capsids that change color depending on the stage of the infectious cycle, we find that during entry, axons of PNS neurons support robust, bidirectional capsid motility, similar to cellular cargo, toward the cell body. In contrast, progeny particles appear to be transported unidirectionally by kinesin motors toward distal egress sites.A lphaherpesviruses (e.g., human herpes simplex virus [HSV] and swine pseudorabies virus [PRV]) are neuroinvasive double-stranded DNA (dsDNA) viruses with virions consisting of a nucleocapsid surrounded by a proteinaceous tegu...

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PKA-dependent dynein switching from lysosomes to adenovirus: A novel form of host–virus competition

Abstract: PKA-mediated phosphorylation of a specific residue in the dynein light intermediate chain 1 releases the motor protein from lysosomes and late endosomes while activating its recruitment to adenovirus capsids.

Cited by 64 publications

References 59 publications

Adenovirus Modulates Toll-Like Receptor 4 Signaling by Reprogramming ORP1L-VAP Protein Contacts for Cholesterol Transport from Endosomes to the Endoplasmic Reticulum

Adenovirus Modulates Toll-Like Receptor 4 Signaling by Reprogramming ORP1L-VAP Protein Contacts for Cholesterol Transport from Endosomes to the Endoplasmic Reticulum

Hepatitis C virus promotes virion secretion through cleavage of the Rab7 adaptor protein RILP

Dual-Color Herpesvirus Capsids Discriminate Inoculum from Progeny and Reveal Axonal Transport Dynamics

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