![\"Jonathan](\"https:\/\/depts.washington.edu\/jaisril\/wordpress\/wp-content\/uploads\/2015\/09\/jon-180x199.png\")
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Jonathan C. Reed is a Senior Research Scientist in the Lingappa Lab.\u00a0 Jon received his BS from Oregon State University (OSU), where he majored in Life Sciences\/Botany and studied suppressors of RNA silencing encoded by plant viruses, in the laboratory of\u00a0Dr. Valerian Dolja.\u00a0 At OSU, Jon received numerous awards, including two\u00a0HHMI summer undergraduate research fellowships\u00a0and the Outstanding Senior Award at Oregon State University.\u00a0 In 2004 Jon entered the University of Washington (UW) Molecular and Cellular Biology PhD program, and in 2006 he received an NSF Graduate Research Fellowship\u00a0to pursue his PhD studies on capsid assembly in the Lingappa lab.\u00a0 \u00a0In 2011 he obtained his PhD in Molecular and Cellular Biology and his MS in Epidemiology through a\u00a0joint UW PhD\/MS program.\u00a0 Jon stayed on in the Lingappa lab as a postdoctoral fellow until 2013, when he joined Prosetta Biosciences<\/a><\/u> and established the Prosetta satellite lab in Seattle.\u00a0 While at Prosetta, Jon served as Biology Lead and directed two different groups \u2013 one screening for anti-viral compounds that act on the subcellular ribonucleoprotein assembly machines<\/em> that Jon studied while he was in the Lingappa lab, and one screening for anti-parasitic compounds that act on similar ribonucleoprotein complexes in the malaria parasite.\u00a0 In 2017, Jon returned to Jaisri Lingappa\u2019s lab at UW as a Senior Research Scientist to continue his studies of antiretroviral compounds in collaboration with Prosetta and to study the basic science of retroviral assembly.<\/p>\n As a PhD student in the Lingappa lab, Jon made a fundamental discovery that reshaped our understanding of HIV-1 virus assembly.\u00a0 He demonstrated that HIV-1 assembly intermediates (aka assembly machines<\/em>), which form during assembly of the HIV-1 immature capsid, are composed of cellular proteins found in RNA granules.\u00a0 RNA granules are important subcellular host ribonucleoprotein complexes involved in all aspects of RNA metabolism except translation, but had not been implicated previously in HIV-1 assembly.\u00a0 Jon also examined the role of these host proteins in HIV-1 assembly using siRNA knockdown approaches.\u00a0 In these studies, he demonstrated that DDX6, one of the host RNA granule proteins he identified in HIV-1 assembly intermediates, facilitates HIV-1 capsid assembly in cells.\u00a0 He further found that WT DDX6 rescued the knockdown phenotype, while an enzymatically inactive DDX6 mutant did not, thereby demonstrating that DDX6 acts enzymatically to promote HIV-1 assembly.\u00a0 Jon confirmed the physiological relevance of his findings with siRNA knockdowns in primary blood mononuclear cells (PBMC), showing that DDX6 facilitates assembly of infectious virus.\u00a0 Jon\u2019s seminal PhD studies, published in the Journal of Cell Biology in 2012, support of a model in which HIV-1 Gag co-opts host RNA granules to form assembly intermediates and uses the host proteins in these complexes to facilitate capsid assembly, thereby adding to growing evidence that HIV-1 capsid assembly in cells is host-catalyzed and energy-dependent.<\/p>\n As a postdoctoral fellow, Jon developed RT-qPCR techniques to allow us to ask whether HIV-1<\/em> assembly machines<\/em>\u00a0also facilitate packaging of the HIV-1 genome into newly assembled capsids. \u00a0These advances led to a paper, published in PLOS Pathogens in 2018, showing that the RNA-granule-derived HIV-1 assembly intermediates contain much of the cytoplasmic HIV-1 genomic RNA found in HIV-expressing cells.\u00a0 Thus, by trafficking to these RNA granules, assembling HIV-1 Gag is able to access host facilitators of assembly (e.g. DDX6 and the host enzyme ABCE1) as well as the HIV-1 genomic RNA that must be encapsidated during assembly for the resulting virus to be infectious.<\/p>\n As a Senior Research Scientist in the Lingappa Lab, Jon demonstrated that the Gag protein of feline immunodeficiency virus (FIV), a non-primate lentivirus, forms RNA-granule-derived assembly intermediates containing ABCE1 and DDX6, analogous to what is observed for HIV-1.\u00a0 These findings suggest that retroviral assembly machines are evolutionarily conserved.\u00a0 This study, published in the Journal of Virology in 2018, also demonstrated that the RNA-granule-derived FIV assembly intermediates contain additional RNA granule proteins, such as DCP-2, but do not contain the abundant ribosomal protein S6.\u00a0 Thus, these complexes resemble RNA granules but do not resemble ribosomes.\u00a0 This paper also features imaging studies in which Jon used the proximity ligation assay to demonstrate colocalization of FIV Gag with RNA granule proteins.<\/p>\n Jon\u2019s most recent project describes a novel small molecule (PAV117), discovered in collaboration with Prosetta Biosciences, that inhibits replication of HIV-1 in T cell lines and primary T cells with nanomolar potency, most likely acting on HIV-1 assembly intermediates (manuscript in preparation).<\/p>\n Jon\u2019s contributions to the Lingappa Lab are myriad, and include writing manuscripts, editing grants, supervising junior employees, helping to plan long-term research goals, generating ideas for new projects, researching the literature, and developing new methods.\u00a0 He has piloted, optimized, and established a vast number of diverse techniques in the Lingappa lab, including highly sensitive coimmunoprecipitation approaches, quantitative immunogold electron microscopic double labeling, quantitative proximity ligation assays for studying subcellular localization of small molecules, siRNA knockdown and rescue, codon optimization, reverse transcription with quantitative PCR, northern blotting, a wide variety of infectivity assays, other virologic assays, and assays for studying inhibition of viral replication by antiviral small molecules including in primary cells.<\/p>\n 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