Gregory Allan Smith
Northwestern University At Chicago
Project start date: 2010-02-01
Project end date: 2015-01-31
Sponsored Links Excellgen http://Excellgen.com
Grants awarded to Gregory Allan Smith
ALPHA-HERPEVIRUS ASSEMBLY EGRESS AND VIRAL PARTICLE HETEROGENEITY
Gregory Allan Smith, Assistant Professor
Northwestern University, 750 N. Lake Shore Drive, 7th, Chicago, Il 60611
Grant 1R01AI080658-01A2 from National Institute Of Allergy And Infectious Diseases
Abstract: The neuroinvasive herpesviruses are a highly-prevalent group of the alpha-herpesvirus subfamily that includes the human pathogens herpes simplex virus types 1 and 2 (HSV-1, HSV-2), and varicella zoster virus (VZV). An additional member of this group is a virus of veterinary significance, pseudorabies virus (PRV), which historically has provided models for studying viral pathogenesis. Despite the availability of the antiviral compound acyclovir, several severe forms of disease caused by these viruses remain prevalent in this country and worldwide. Infections associated with high rates of morbidity or mortality includes encephalitis, keratitis, shingles and disseminated infections in newborns. Novel strategies to interfere with the assembly and egress of these viruses could prove valuable to treatment of infections, yet much of the herpesvirus infectious cycle remains undefined. In this application we leverage our expertise in infectious clone mutagenesis and single viral particle fluorescence imaging methods to dissect viral structural composition in living-cells and extracellularly, and to address the molecular pathways guiding viral assembly and egress. New evidence is provided indicating that the very large herpesvirus tegument protein, VP1/2, is a key effector of viral assembly during distinct stages of assembly in the nucleus and cytoplasm of infected cells. This proposal is based on the hypothesis that herpesvirus assembly and egress are coupled processes that occur through a series of sequential steps both in the nucleus and cytoplasm of infected cells, and that each of these steps are affected in part by the VP1/2 protein. Our goal is to refine our understanding of these steps at the level of the protein interactions that contribute to the dynamics of viral egress. This proposal includes comparative studies of model viruses from the two neuroinvasive herpesviruses subgroups, the simplex viruses (represented by HSV-1) and the varicelloviruses (represented by PRV), to develop a comprehensive analysis of the properties of these viruses. Neuroinvasive herpesviruses are the causative agents of a number of severe diseases including shingles, encephalitis, neonatal infections and herpes keratitis (the leading cause of infectious blindness in the USA and other industrialized nations). This proposal focuses on understanding the molecular mechanisms that underlie the assembly and egress of herpesvirus particles, with the long- term goal of identifying new targets for the intervention of disease progression
Keywords: 0-6 weeks old; 2-Amino-1, 9-dihydro-9-[(2-hydroxyethoxy)methyl]-6H-purin-6-one; 9-((2-Hydroxyethoxy)methyl)guanine; Acicloftal; Aclovir; Acycloguanosine; Acyclovir; Address; Affect; Antiviral Agents; Antiviral Drugs; Antivirals; Aujeszky`s Disease Virus; Aujeszkys Disease Virus; Blindness; Capsid; Capsid Proteins; Cargosil; Cell Nucleus; Cells; Chickenpox Virus; Coat Proteins; Comparative Study; Country; Coupled; Crystallography, X-Ray; Crystallography, X-Ray Diffraction; Crystallography, X-Ray/Neutron; Crystallography, Xray; Cytoplasm; Developed Countries; Developed Nations; Disease; Disease Progression; Disorder; Encephalitis; Genetics-Mutagenesis; Glaxo Wellcome Brand of Aciclovir; Glaxo Wellcome Brand of Aciclovir Sodium Salt; Goals; HHV-1; HHV-2; HHV-3; HSV; HSV-1; HSV-2; HSV1; HSV2; Herpes Simplex Virus; Herpes Simplex Virus 1; Herpes Simplex Virus 2; Herpes Simplex Virus Type 1; Herpes Simplex Virus Type 2; Herpes Zoster; Herpes labialis Virus; Herpes zoster Virus; Herpes zoster disease; Herpesviridae; Herpesvirus 1 (alpha), Human; Herpesvirus 1 (alpha), Suid; Herpesvirus 1, Human; Herpesvirus 1, Suid; Herpesvirus 2 (alpha), Human; Herpesvirus 2, Human; Herpesvirus 3 (alpha), Human; Herpesvirus Suis; Herpesvirus hominis; Herpesvirus progenitalis; Herpesvirus varicellae; Herpesviruses; Heterogeneity; Human; Human (alpha) herpes virus 2; Human Herpesvirus 2; Human herpes simplex virus type 1; Human herpes simplex virus type 2; Human herpesvirus 1; Human herpesvirus 3; Human herpesvirus type 1; Human, General; Individual; Industrialized Countries; Industrialized Nations; Infant, Newborn; Infection; Inflammation, Brain; Intervention; Intervention Strategies; Keratitis; Life; Link; Man (Taxonomy); Man, Modern; Measures; Molecular; Molecular Biology, Mutagenesis; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Mutagenesis; Neonatal; Newborn Infant; Newborns; Nuclear; Nuclear Envelope; Nuclear Membrane; Nucleus; Ocular Herpes zoster Virus; Parke Davis Brand of Aciclovir; Pathway interactions; Poviral; Process; Property; Property, LOINC Axis 2; Proteins; Pseudorabies virus; Role; SUBGP; Series; Shingles; Simplexvirus; Single Crystal Diffraction; Staging; Structure; Study models; Subgroup; Suid Herpesvirus 1; Swine Herpesvirus 1; VIVIPAROUS1 protein; VZ Virus; Varicella-Zoster Virus; Varicellovirus; Viral; Viral Coat Proteins; Viral Diseases; Viral Gene Products; Viral Gene Proteins; Viral Outer Coat Protein; Viral Pathogenesis; Viral Proteins; Virorax; Virus; Virus Diseases; Viruses, General; Viviparous-1 protein; Vp1 protein; Warner Wellcome Brand of Aciclovir; Wellcome Brand of Aciclovir; Wellcome Brand of Aciclovir Sodium Salt; X Ray Crystallographies; X-Ray Crystallography; Zona; Zoster; Zovirax; Zovirax for Injection; base; coat (nveloped virus); disease/disorder; domain mapping; fluorescence imaging; gene product; herpes simplex i; herpes simplex ii; herpes virus; herpes virus 1, human; herpes zona; herpesvirus; human alphaherpesvirus 1; human alphaherpesvirus 2; human alphaherpesvirus 3; human herpesvirus 1 group; human herpesvirus 3 group; imaging modality; interventional strategy; member; mutant; new approaches; newborn human (0-6 weeks); novel approaches; novel strategies; novel strategy; particle; pathogen; pathway; public health relevance; social role; viral infection; virus infection; virus protein
Relevance: Neuroinvasive herpesviruses are the causative agents of a number of severe diseases including shingles, encephalitis, neonatal infections and herpes keratitis (the leading cause of infectious blindness in the USA and other industrialized nations). This proposal focuses on understanding the molecular mechanisms that underlie the assembly and egress of herpesvirus particles, with the long- term goal of identifying new targets for the intervention of disease progression
Project start date: 2010-02-01
Project end date: 2015-01-31
Budget start date: 1-FEB-2010
Budget end date: 31-JAN-2011
PFA/PA: PA-07-070
1R01AI080658-01A2 (2010): $305000
ALPHA-HERPESVIRUS TRANSPORT ON AXONS
Gregory Allan Smith, Assistant Professor
Northwestern University, 750 N. Lake Shore Drive, 7th, Chicago, Il 60611
Grant 2R01AI056346-06A2 from National Institute Of Allergy And Infectious Diseases
Abstract: Spread of neuroinvasive herpesviruses from sensory neurons to the eye, brain or from mother to newborn are leading causes of morbidity and mortality associated with infections by this class of pathogens. Herpes simplex virus type 1 (HSV-1) and pseudorabies virus (PRV) are representative members of the two genuses of neuroinvasive herpesviruses (simplexviruses & varicelloviruses), both belonging to the alpha-herpesvirus subfamily. Both viruses establish life-long latent infections in sensory neurons of the peripheral nervous system, and both are established models for use in the laboratory. In this proposal, we leverage our strengths in infectious clone mutagenesis and live-cell viral tracking methods to address the mechanisms of HSV-1 and PRV spread within sensory neurons. Viral transport to the site of latency and transport following reactivation to peripheral innervated tissues are both critical to the viral infectious cycle, and both of these stages of infection are modeled using cultured sensory neurons and examined in animals to investigate the mechanisms of virus intracellular trafficking. In addition, new evidence is provided indicating that the very large herpesvirus tegument protein, VP1/2, is responsible for intracellular transport of capsids, and new tools for dissecting the mechanism of viral transport along microtubules are employed. Neuroinvasive alpha-herpesviruses are the causative agents of a number of severe diseases including shingles, encephalitis, neonatal infections and herpes keratitis (the leading cause of infectious blindness in the USA and other industrialized nations). This proposal focuses on understanding the cellular mechanisms used by herpesviruses to spread within the nervous system and cause disease, with the long term goal of developing new treatments to intervene with disease progression
Keywords: 0-6 weeks old; Address; Afferent Neurons; Animal Model; Animal Models and Related Studies; Animals; Assay; Aujeszky`s Disease Virus; Aujeszkys Disease Virus; Axon; Axon Terminals; Axonal Transport; Axoplasmic Transport; Bioassay; Biologic Assays; Biological Assay; Blindness; Body Tissues; Brain; Capsid; Cells; Central Nervous System; Chickenpox Virus; Complex; Defect; Developed Countries; Developed Nations; Disease; Disease Progression; Disorder; Electron Microscopy; Encephalitis; Encephalon; Encephalons; Envelope Protein; Event; Eye; Eyeball; Fluorescence; Funding; Gene Products, env; Genetics-Mutagenesis; Goals; HHV-1; HHV-2; HHV-3; HSV; HSV-1; HSV-2; HSV1; HSV2; Herpes Simplex Virus; Herpes Simplex Virus 1; Herpes Simplex Virus 2; Herpes Simplex Virus Type 1; Herpes Simplex Virus Type 2; Herpes Zoster; Herpes labialis Virus; Herpes zoster Virus; Herpes zoster disease; Herpesviridae; Herpesvirus 1 (alpha), Human; Herpesvirus 1 (alpha), Suid; Herpesvirus 1, Human; Herpesvirus 1, Suid; Herpesvirus 2 (alpha), Human; Herpesvirus 2, Human; Herpesvirus 3 (alpha), Human; Herpesvirus Suis; Herpesvirus hominis; Herpesvirus progenitalis; Herpesvirus varicellae; Herpesviruses; Human; Human (alpha) herpes virus 2; Human Herpesvirus 2; Human herpes simplex virus type 1; Human herpes simplex virus type 2; Human herpesvirus 1; Human herpesvirus 3; Human herpesvirus type 1; Human, General; Image; Industrialized Countries; Industrialized Nations; Infant, Newborn; Infection; Inflammation, Brain; Intracellular Transport; Keratitis; Kinesin; Kinetic; Kinetics; Laboratories; Life; Link; Man (Taxonomy); Man, Modern; Maps; Measures; Methods; Micro-tubule; Microtubules; Modeling; Molecular; Molecular Biology, Mutagenesis; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Mothers; Motor; Mutagenesis; NRVS-SYS; Neonatal; Nerve Cells; Nerve Endings, Presynaptic; Nerve Unit; Nervous System; Nervous System, Brain; Nervous System, CNS; Nervous system structure; Neural Cell; Neural Transmission; Neuraxis; Neurocyte; Neurologic Body System; Neurologic Organ System; Neurons; Neurons, Afferent; Neurons, Sensory; Newborn Infant; Newborns; Ocular Herpes zoster Virus; Pathway interactions; Peripheral; Peripheral Nerves; Peripheral Nervous System; Phase; Presynaptic Terminals; Property; Property, LOINC Axis 2; Proteins; Pseudorabies virus; Recombinants; Recruitment Activity; Role; Secretory Component; Secretory Piece; Sensory Cell Afferent Neuron; Shingles; Simplexvirus; Site; Staging; Study models; Suid Herpesvirus 1; Swine Herpesvirus 1; Synaptic Boutons; Synaptic Terminals; Synaptic Transmission; System; System, LOINC Axis 4; Testing; Time; Tissues; Transmission; Transport Piece; VIVIPAROUS1 protein; VZ Virus; Varicella-Zoster Virus; Varicellovirus; Viral; Viral Pathogenesis; Virion; Virus; Virus Particle; Viruses, General; Viviparous-1 protein; Vp1 protein; Work; Zona; Zoster; coat (nveloped virus); design; designing; disease/disorder; env Antigens; env Gene Products; env Polyproteins; env Protein; experiment; experimental research; experimental study; fluorescence imaging; gene product; herpes simplex i; herpes simplex ii; herpes virus; herpes virus 1, human; herpes zona; herpesvirus; human alphaherpesvirus 1; human alphaherpesvirus 2; human alphaherpesvirus 3; human herpesvirus 1 group; human herpesvirus 3 group; imaging; latent infection; member; model organism; mutant; neuronal; newborn human (0-6 weeks); particle; pathogen; pathway; public health relevance; recruit; research study; social role; tomography; tool; trafficking; transmission process
Relevance: Neuroinvasive alpha-herpesviruses are the causative agents of a number of severe diseases including shingles, encephalitis, neonatal infections and herpes keratitis (the leading cause of infectious blindness in the USA and other industrialized nations). This proposal focuses on understanding the cellular mechanisms used by herpesviruses to spread within the nervous system and cause disease, with the long term goal of developing new treatments to intervene with disease progression
Project start date: 2003-07-01
Project end date: 2014-11-30
Budget start date: 15-DEC-2009
Budget end date: 30-NOV-2010
PFA/PA: PA-07-070
2R01AI056346-06A2 (2010): $186813
Alpha-Herpesvirus Transport In Axons
Gregory Allan Smith
Microbiology-immunologynorthwestern University
750 N. Lake Shore Drive, 7th
chicago, Il 60611
Grant 5R01AI056346-05 from National Institute Of Allergy And Infectious Diseases IRG: VR
Abstract: Alpha-herpesviruses are neurotropic pathogens that include the human viruses varicella-zoster virus (VZV) and herpes simplex virus (HSV) types 1 and 2, and the animal virus pseudorabies virus (PRV). These viruses spread into the nervous system by directed axonal transport to sensory neuron cell bodies in peripheral ganglia, where life-long latent infections are established. Infections are typically not life threatening in healthy individuals, however, these viruses have the capacity to spread trans-synaptically to enter the brain by circuit-specific routes. Once in the brain, the resulting encephalitis is lethal in the majority of cases. The long-term goals of my studies are to determine the mechanisms by which alpha-herpesviruses spread in the nervous system, and to identify the factors governing disease outcome. This application will focus on virus spread in the axons of sensory neurons. Specifically Using infectious clone mutagenesis methods, viruses will be isolated carrying mutations in the genes encoding each tegument protein. The mutant viruses will be screened for defects in capsid transport in axons by time-lapse tracking of capsids fused to the green-fluorescence protein. Structure/function analysis of tegument proteins required for capsid transport will be initiated. In this way, the role of the viral tegument proteins in capsid transport will be definitively tested. These proteins are expected to both tether capsids to microtubule motors and regulate motor activity, as no other class of viral proteins are known to associate with capsids and be exposed to the host cytosol simultaneously. The results of these studies will determine how alpha-herpesviruses move directionally within sensory neurons. This will serve as a first step to identifying the molecular mechanisms of viral transport, and to addressing the larger question of virus spread in the vertebrate nervous system
Keywords: Alphaherpesvirinae, capsid, neuronal transport, protein structure function, virus infection mechanism, virus protein afferent nerve, central neural pathway /tract, encephalitis, gene mutation, host organism interaction, latent virus infection, microtubule associated protein, neuroregulation, recombinant virus artificial chromosome, fluorescence microscopy, genetic screening, green fluorescent protein, site directed mutagenesis, tissue /cell culture
Project start date: 2004-02-01
Project end date: 2010-01-31
5R01AI056346-05 (2008): $276259
5R01AI056346-04 (2007): $281609
5R01AI056346-03 (2006): $290021
5R01AI056346-02 (2005): $297000
1R01AI056346-01A1 (2004): $297000
3R01AI056346-01A1S1 (2004): $8787
Gregory Allan Smith
Northwestern University At Chicago
Project start date: 2003-07-01
Project end date: 2014-11-30
ALPHA-HERPESVIRUS TRANSPORT ON AXONS
Gregory Allan Smith
Department/ Educational Institution Type:
Grant 5R01AI056346-07 from National Institute Of Allergy And Infectious Diseases
Keywords: 0-6 weeks old; Address; Afferent Neurons; Animal Model; Animal Models and Related Studies; Animals; Assay; Aujeszky`s Disease Virus; Aujeszkys Disease Virus; Axon; Axon Terminals; Axonal Transport; Axoplasmic Transport; Bioassay; Biologic Assays; Biological Assay; Blindness; Body Tissues; Brain; Capsid; Cells; Central Nervous System; Chickenpox Virus; coat (nveloped virus); Complex; Defect; design; designing; Developed Countries; Developed Nations; Disease; Disease Progression; disease/disorder; Disorder; Electron Microscopy; Encephalitis; Encephalon; Encephalons; env Antigens; env Gene Products; env Polyproteins; env Protein; Envelope Protein; Event; experiment; experimental research; experimental study; Eye; Eyeball; Fluorescence; fluorescence imaging; Funding; gene product; Gene Products, env; Genetics-Mutagenesis; Goals; Herpes labialis Virus; herpes simplex i; herpes simplex ii; Herpes Simplex Virus; Herpes Simplex Virus 1; Herpes Simplex Virus 2; Herpes Simplex Virus Type 1; Herpes Simplex Virus Type 2; herpes virus; herpes virus 1, human; herpes zona; Herpes Zoster; Herpes zoster disease; Herpes zoster Virus; Herpesviridae; herpesvirus; Herpesvirus 1 (alpha), Human; Herpesvirus 1 (alpha), Suid; Herpesvirus 1, Human; Herpesvirus 1, Suid; Herpesvirus 2 (alpha), Human; Herpesvirus 2, Human; Herpesvirus 3 (alpha), Human; Herpesvirus hominis; Herpesvirus progenitalis; Herpesvirus Suis; Herpesvirus varicellae; Herpesviruses; HHV-1; HHV-2; HHV-3; HSV; HSV-1; HSV-2; HSV1; HSV2; Human; Human (alpha) herpes virus 2; human alphaherpesvirus 1; human alphaherpesvirus 2; human alphaherpesvirus 3; Human herpes simplex virus type 1; Human herpes simplex virus type 2; Human herpesvirus 1; human herpesvirus 1 group; Human Herpesvirus 2; Human herpesvirus 3; human herpesvirus 3 group; Human herpesvirus type 1; Human, General; Image; imaging; Industrialized Countries; Industrialized Nations; Infant, Newborn; Infection; Inflammation, Brain; Intracellular Transport; Keratitis; Kinesin; Kinetic; Kinetics; Laboratories; latent infection; Life; Link; Man (Taxonomy); Man, Modern; Maps; Measures; member; Methods; Micro-tubule; Microtubules; model organism; Modeling; Molecular; Molecular Biology, Mutagenesis; Morbidity; Morbidity - disease rate; Mortality; Mortality Vital Statistics; Mothers; Motor; Mutagenesis; mutant; Neonatal; Nerve Cells; Nerve Endings, Presynaptic; Nerve Unit; Nervous System; Nervous system structure; Nervous System, Brain; Nervous System, CNS; Neural Cell; Neural Transmission; Neuraxis; Neurocyte; Neurologic Body System; Neurologic Organ System; neuronal; Neurons; Neurons, Afferent; Neurons, Sensory; newborn human (0-6 weeks); Newborn Infant; Newborns; NRVS-SYS; Ocular Herpes zoster Virus; particle; pathogen; pathway; Pathway interactions; Peripheral; Peripheral Nerves; Peripheral Nervous System; Phase; Presynaptic Terminals; Property; Property, LOINC Axis 2; Proteins; Pseudorabies virus; public health relevance; Recombinants; recruit; Recruitment Activity; research study; Role; Secretory Component; Secretory Piece; Sensory Cell Afferent Neuron; Shingles; Simplexvirus; Site; social role; Staging; Study models; Suid Herpesvirus 1; Swine Herpesvirus 1; Synaptic Boutons; Synaptic Terminals; Synaptic Transmission; System; System, LOINC Axis 4; Testing; Time; Tissues; tomography; tool; trafficking; Transmission; transmission process; Transport Piece; Varicella-Zoster Virus; Varicellovirus; Viral; Viral Pathogenesis; Virion; Virus; Virus Particle; Viruses, General; Viviparous-1 protein; VIVIPAROUS1 protein; Vp1 protein; VZ Virus; Work; Zona; Zoster
Relevance: Neuroinvasive alpha-herpesviruses are the causative agents of a number of severe diseases including shingles, encephalitis, neonatal infections and herpes keratitis (the leading cause of infectious blindness in the USA and other industrialized nations). This proposal focuses on understanding the cellular mechanisms used by herpesviruses to spread within the nervous system and cause disease, with the long term goal of developing new treatments to intervene with disease progression
Project start date: 2003-07-01
Project end date: 2014-11-30
Budget start date: 1-DEC-2010
Budget end date: 30-NOV-2011
PFA/PA: PA-07-070
5R01AI056346-07 (2011): $308813
Sponsored Links Excellgen http://Excellgen.com