Network of Interactions between ZIKA Virus Non-Structural Proteins and Human Host Proteins

Abstract

The Zika virus (ZIKV) is a mosquito-borne Flavivirus and can be transmitted through an infected mosquito bite or through human-to-human interaction by sexual activity, blood transfusion, breastfeeding, or perinatal exposure. After the 2015–2016 outbreak in Brazil, a strong link between ZIKV infection and microcephaly emerged. ZIKV specifically targets human neural progenitor cells, suggesting that proteins encoded by ZIKV bind and inactivate host cell proteins, leading to microcephaly. Here, we present a systematic annotation of interactions between human proteins and the seven non-structural ZIKV proteins corresponding to a Brazilian isolate. The interaction network was generated by combining tandem-affinity purification followed by mass spectrometry with yeast two-hybrid screens. We identified 150 human proteins, involved in distinct biological processes, as interactors to ZIKV non-structural proteins. Our interacting network is composed of proteins that have been previously associated with microcephaly in human genetic disorders and/or animal models. Further, we show that the protein inhibitor of activated STAT1 (PIAS1) interacts with NS5 and modulates its stability. This study builds on previously published interacting networks of ZIKV and genes related to autosomal recessive primary microcephaly to generate a catalog of human cellular targets of ZIKV proteins implicated in processes related to microcephaly in humans. Collectively, these data can be used as a resource for future characterization of ZIKV infection biology and help create a basis for the discovery of drugs that may disrupt the interaction and reduce the health damage to the fetus.

Inter-organelle interactions between the ER and mitotic spindle facilitates Zika protease cleavage of human Kinesin-5 and results in mitotic defects

“…Prior studies designed to identify and confirm cellular targets of Zika protease commonly utilize a well-characterized chimera fusion between a soluble loop from NS2B transmembrane component and the NS3 protease domain that expresses as a soluble active protease in cells (Yamaoka et al, 2021;De Jesú s-Gonzá lez et al, 2020;Li et al, 2019;Hill et al, 2018;Coyaud et al, 2018;Shah et al, 2018;Scaturro et al, 2018). More recent proteomic analyses expressed the NS3 component in the absence of the NS2B subunit (Golubeva et al, 2020;Coyaud et al, 2018). In our hands, the NS2B-NS3 chimera, s-ZPrC, exhibited toxicity in cells and cell death of the majority of transfected cells, similar to that observed in previous reports (not shown) (Arias-Arias et al, 2020;Li et al, 2019).…”

“…Viral proteins directly interact with cellular components, via protein-protein interactions, to exert this control. In the case of Zika, with its limited proteome, the NS2B-NS3 protease is known to cleave host proteins (Golubeva et al, 2020;Hill et al, 2018;Coyaud et al, 2018;Shah et al, 2018;Scaturro et al, 2018), whereas both the NS4 and NS5 proteins collaborate to inhibit the establishment of cellular antiviral state by blocking the interferon (IFN)-a/b pathway (Liang et al, 2016;Dar et al, 2017;Grant et al, 2016;Kumar et al, 2016). Numerous recent studies have documented that Zika virus can impair neurogenesis and interfere with the proliferation of neuronal precursor cells (Ferreira and Garcez, natively ER membrane bound by NS2B, but with a C-terminal truncation in NS3 that removes the helicase domain.…”

Comprehensive host-pathogen protein-protein interaction network analysis

“…There are many real bipartite networks such as actor-movie network [35] which illustrates the movies and the actors playing in them, author-article network [36] which shows the articles and theirs authors, gene-disease network [37][38][39][40][41][42][43] which reveals diseases and their targeted genes, metabolite-enzyme network [44][45][46] which shows metabolites and their corresponding enzymes, and host-pathogen PPI network such as fungal pathogen scedosporium aurantiacum with human long epithelial cells [47], Leptospira interrogans and Homo sapiens [48], extracellular bacterial pathogen and human host [49], zika virus non-structural proteins and human host proteins [50], bacterial and fungal pathogens and maize stalks host [51].…”

The journey of Zika to the developing brain

“…These pathways may be crucial in understanding ZIKV infection and subsequent microcephaly in foetuses. Recent proteomics studies exploring the ZIKV-host interactome have identified, amongst an extensive array of cellular targets, networks associated with signalling and mitotic pathways [27][28][29][30]. For example, Golubeva et al identify shared interactions with 14 of the reported MCPH-related genes [30].…”