Autophagy plays a paramount role in mammalian antiviral immunity including direct targeting of viruses and their individual components, and many viruses have evolved measures to antagonize or even exploit autophagy mechanisms for the benefit of infection. In plants, however, the functions of autophagy in host immunity and viral pathogenesis are poorly understood. In this study, we have identified both anti-and proviral roles of autophagy in the compatible interaction of cauliflower mosaic virus (CaMV), a double-stranded DNA pararetrovirus, with the model plant Arabidopsis thaliana. We show that the autophagy cargo receptor NEIGHBOR OF BRCA1 (NBR1) targets nonassembled and virus particle-forming capsid proteins to mediate their autophagy-dependent degradation, thereby restricting the establishment of CaMV infection. Intriguingly, the CaMV-induced virus factory inclusions seem to protect against autophagic destruction by sequestering capsid proteins and coordinating particle assembly and storage. In addition, we found that virus-triggered autophagy prevents extensive senescence and tissue death of infected plants in a largely NBR1-independent manner. This survival function significantly extends the timespan of virus production, thereby increasing the chances for virus particle acquisition by aphid vectors and CaMV transmission. Together, our results provide evidence for the integration of selective autophagy into plant immunity against viruses and reveal potential viral strategies to evade and adapt autophagic processes for successful pathogenesis. A utophagy is a conserved intracellular pathway that engages specialized double-membrane vesicles, called "autophagosomes," to enclose and transport cytoplasmic content to lytic compartments for degradation and subsequent recycling (1). Autophagosome formation relies on extensive membrane rearrangements and is mediated by the concerted action of a core set of autophagy-related proteins (ATGs) (2, 3). At basal levels, autophagy serves mainly housekeeping functions in cellular homeostasis, whereas stimulated autophagy activity facilitates adaptation to developmental and environmental stress conditions including starvation, aging, and pathogen infection (1, 4). Ample evidence now indicates that autophagy, initially recognized as a mainly bulk catabolic process, is able specifically to target and degrade a multitude of cellular structures ranging from individual and aggregated proteins to entire organelles and invading microbes (5, 6). Selectivity is provided by a growing number of autophagic adaptor or receptor proteins identified in eukaryotic organisms that recruit the cargo to the developing autophagosome through interaction with membrane-associated ATG8/LC3 proteins (7, 8). Several mammalian autophagy receptors have been implicated in the targeting of intracellular bacterial and viral pathogens in a process called "xenophagy" (8-10). For instance, the cargo receptor p62 (SQSTM1) was shown to bind directly to and mediate autophagic clearance of different viral capsid pr...

The helper component proteinase (HC-Pro) is a key protein encoded by plant viruses of the genus Potyvirus. HC-Pro is involved in different steps of the viral cycle, aphid transmission, replication, and virus cell-to-cell and systemic movement and is a suppressor of posttranscriptional gene silencing. Structural knowledge of HC-Pro is required to better understand its multiple functions. To this aim, we purified His-tagged wild-type HC-Pro and a N-terminal deletion mutant (⌬HC-Pro) from plants infected with recombinant potyviruses. Biochemical analysis of the recombinant proteins confirmed that HC-Pro is a dimer in solution, that the N terminus is not essential for self-interaction, and that a large C-terminal domain is highly resistant to proteolysis. Two-dimensional crystals of the recombinant proteins were successfully grown on Ni 2؉ -chelating lipid monolayers. Comparison of projection maps of negatively stained crystals revealed that HC-Pro is composed of two domains separated by a flexible constriction. Cryo-electron crystallography of ⌬HC-Pro allowed us to calculate a projection map at 9-Å resolution. Our data from electron microscopy, biochemical analysis, and secondary structure predictions lead us to suggest a model for structure/function relationships in the HCPro protein.

aphid ͉ receptor N early all plant viruses that cause extensive agricultural damage use specific vectors to spread between hosts. The most common vectors are arthropods, especially aphids (1), and the most widely adopted strategy for virus-vector interaction is noncirculative transmission, in which the virus is taken up by a vector feeding on an infected plant, adsorbed somewhere on the cuticle lining the inner part of the feeding apparatus, and subsequently released to inoculate a new host plant. The viral components involved in this interaction are relatively well established, in particular for the genus Cucumovirus, where domains of the coat protein directly recognize unknown retention sites in the vector mouthparts (capsid strategy), and for the genera Potyvirus and Caulimovirus, where a nonstructural viral protein, HC (helper component), creates the link between virion and vector (helper strategy) (reviewed in refs. 2 and 3). However, no putative binding sites for viral components in the insect vector have ever been chemically characterized or even precisely localized. This question is of major importance, because numerous noncirculative viruses may use the same vector attachment sites, and identification of putative receptor molecules could lead to new strategies to combat viral spread.A distinguishing feature of noncirculative transmission is that several virus species can be transmitted by the same vector, and, conversely, several vector species can transmit the same virus. Hence, although some degree of noncirculative virus/vector specificity exists (4, 5), it is often so broad that the very existence of actual viral receptors remains questionable, because their existence has never been directly demonstrated.Here we report evidence for the existence, precise location, and chemical nature of the receptor for a noncirculative virus, cauliflower mosaic virus (CaMV), in its insect vector. A novel in vitro system allowed rapid visualization of the interaction between dissected aphid stylets and the HC of CaMV. The CaMV retention sites are concentrated exclusively in a tiny and previously unknown anatomical zone located at the extreme tip of the aphid maxillary stylets. Virus/vector binding at this specific zone is mandatory for successful CaMV transmission. Pretreatment of dissected stylets with various chemicals and enzymes demonstrated that the molecule used by CaMV as a specific receptor for vector transmission is a nonglycosylated protein deeply embedded in the chitin matrix.