title: Quantitative proteomic analysis of HIV-1 Tat-induced dysregulation in SH-SY5Y neuroblastoma cells.
abstract: Despite affecting up to 70% of HIV-positive patients and being the leading cause of dementia in patients under 40 years, the molecular mechanisms involved in the onset of HIV-associated neurocognitive disorders (HAND) are not well understood. To address this, we performed SILAC-based quantitative proteomic analysis on HIV-Tat treated SH-SY5Y neuroblastoma cells. Isolated protein was fractionated by SDS-PAGE and analyzed by nLC-MS/MS on an Orbitrap Velos. Using MaxQuant, we identified and quantified 3077 unique protein groups, of which 407 were differentially regulated. After applying an additional standard deviation-based cutoff, 29 of these were identified as highly significantly and stably dysregulated. GO term analysis shows dysregulation in both protein translation machinery as well as cytoskeletal regulation that have both been implicated in other dementias. In addition, several key cytoskeletal regulatory proteins such as ARHGEF17, the Rho GTPase, SHROOM3, and CMRP1 are downregulated. Together, these data demonstrate that HIV-Tat can dysregulate neuronal cytoskeletal regulatory proteins that could lead to the major HAND clinical manifestation-synapse loss.
authors.affiliation: Department of Integrative Biomedical Sciences, University of Cape Town, South Africa.
authors.forename: Tariq
authors.initials: T
authors.lastname: Ganief
authors.affiliation: Department of Integrative Biomedical Sciences, University of Cape Town, South Africa.
authors.forename: Putuma
authors.initials: P
authors.lastname: Gqamana
authors.affiliation: Department of Integrative Biomedical Sciences, University of Cape Town, South Africa.
authors.forename: Shaun
authors.initials: S
authors.lastname: Garnett
authors.affiliation: Department of Psychiatry, University of Cape Town, South Africa.
authors.forename: Jackie
authors.initials: J
authors.lastname: Hoare
authors.affiliation: Department of Psychiatry, University of Cape Town, South Africa.|MRC Unit on Anxiety and Stress Disorders, University of Cape Town, South Africa.
authors.forename: Dan J
authors.initials: DJ
authors.lastname: Stein
authors.affiliation: Department of Psychiatry, University of Cape Town, South Africa.
authors.forename: John
authors.initials: J
authors.lastname: Joska
authors.affiliation: Department of Integrative Biomedical Sciences, University of Cape Town, South Africa.
authors.forename: Nelson
authors.initials: N
authors.lastname: Soares
authors.affiliation: Department of Integrative Biomedical Sciences, University of Cape Town, South Africa.|Institute of Infectious Disease and Molecular Medicine, University of Cape Town, South Africa.
authors.forename: Jonathan M
authors.initials: JM
authors.lastname: Blackburn
mesh_terms: D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D063990:Gene Ontology; D015497:HIV-1 / Q000737:chemistry*; D006801:Humans; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology*; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000494:pharmacology*
chemical_list: D020543:Proteome; D054322:tat Gene Products, Human Immunodeficiency Virus
keywords: Cytoskeleton; HIV dementia; Neurodegeneration; Pathway analysis; SILAC