This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.Cryo-electron microscopy (cryo-EM) combined with three-dimensional image reconstruction techniques and newly developed time-resolved methods provide a powerful way to examine virus-host cell interactions as well as the dynamic structural changes that occur during viral cell entry. In fact these methods often represent the only feasible structural approach for large (100 million Dalton) macromolecular complexes and virus/receptor complexes with highly mobile receptor binding sites, as has been postulated for adenovirus. These proposed studies will elucidate the structural events during adenovirus/receptor binding at thle cell surface, intemalization into coatedpits, and escape ofthe virus particle from the endosome.
Specific aims i nclude imaging the adenovirus particle complexed with a Fab fragment of a monoclonal antibody that blocks cell entry in order to localize the Arg-Gly-Asp integrin binding site on the penton base viral component. In addition, the structure of the virus complex ed with a soluble form of the internalization receptor, the integrins avb3 and avb5, will be solved in order to evaluate possible structural changes that occur upon receptor binding. We will also examine conformational changes in the virus and virus/receptor complexes as a consequence of varying the pH value to mimic both the extracellular (pH 8) and early endosomal (pH 6.0 - 6.5) environments. Rapid freezing (within milliseconds) after low pH treatment allows the trapping of structural intermediates. A further objective involves calculating an image reconstruction of the adenovirus temperature sensitive mutant, tsl, which is defective for cell entry. The structural differences between the tsl mutant and wild type will be analyzed to isolate elements essential for productive adenoviral cell entry. These cryo-EM structural studies will provide a greater understanding of the cell entry process of adenovirus, a promising vector for in vivo gene therapy.
| Green, Shulamite A; Hernandez, Leanna M; Bowman, Hilary C et al. (2018) Sensory over-responsivity and social cognition in ASD: Effects of aversive sensory stimuli and attentional modulation on neural responses to social cues. Dev Cogn Neurosci 29:127-139 |
| Yang, Yaling; Joshi, Shantanu H; Jahanshad, Neda et al. (2017) Neural correlates of proactive and reactive aggression in adolescent twins. Aggress Behav 43:230-240 |
| Dennis, Emily L; Rashid, Faisal; Faskowitz, Josh et al. (2017) MAPPING AGE EFFECTS ALONG FIBER TRACTS IN YOUNG ADULTS. Proc IEEE Int Symp Biomed Imaging 2017:101-104 |
| Walsh, Christine M; Ruoff, Leslie; Walker, Kathleen et al. (2017) Sleepless Night and Day, the Plight of Progressive Supranuclear Palsy. Sleep 40: |
| Green, Shulamite A; Hernandez, Leanna; Bookheimer, Susan Y et al. (2017) Reduced modulation of thalamocortical connectivity during exposure to sensory stimuli in ASD. Autism Res 10:801-809 |
| Kodumuri, Nishanth; Sebastian, Rajani; Davis, Cameron et al. (2016) The association of insular stroke with lesion volume. Neuroimage Clin 11:41-45 |
| Kamins, Joshua; Giza, Christopher C (2016) Concussion-Mild Traumatic Brain Injury: Recoverable Injury with Potential for Serious Sequelae. Neurosurg Clin N Am 27:441-52 |
| Agis, Daniel; Goggins, Maria B; Oishi, Kumiko et al. (2016) Picturing the Size and Site of Stroke With an Expanded National Institutes of Health Stroke Scale. Stroke 47:1459-65 |
| Levine, Andrew J; Soontornniyomkij, Virawudh; Achim, Cristian L et al. (2016) Multilevel analysis of neuropathogenesis of neurocognitive impairment in HIV. J Neurovirol 22:431-41 |
| Flournoy, John C; Pfeifer, Jennifer H; Moore, William E et al. (2016) Neural Reactivity to Emotional Faces May Mediate the Relationship Between Childhood Empathy and Adolescent Prosocial Behavior. Child Dev 87:1691-1702 |
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