Title | Inhibition of p66ShcA longevity gene rescues podocytes from HIV-1-induced oxidative stress and apoptosis. |
Publication Type | Journal Article |
Year of Publication | 2009 |
Authors | Husain M, Meggs LG, Vashistha H, Simoes S, Griffiths KO, Kumar D, Mikulak J, Mathieson PW, Saleem MA, Del Valle L, Pina-Oviedo S, Wang JYing, Seshan SV, Malhotra A, Reiss K, Singhal PC |
Journal | J Biol Chem |
Volume | 284 |
Issue | 24 |
Pagination | 16648-16658 |
Date Published | 2009 Jun 12 |
ISSN | 0021-9258 |
Keywords | Active Transport, Cell Nucleus, Apoptosis, Carrier Proteins, Cell Adhesion Molecules, Neuronal, Cell Line, Transformed, Forkhead Box Protein O3, Forkhead Transcription Factors, Green Fluorescent Proteins, HIV Infections, HIV-1, Humans, Membrane Proteins, Oxidative Stress, Phenotype, Phosphorylation, Podocytes, Reactive Oxygen Species, RNA, Small Interfering, Shc Signaling Adaptor Proteins, Src Homology 2 Domain-Containing, Transforming Protein 1, Threonine, Transfection |
Abstract | Glomerular visceral epithelial cells (podocytes) play a critical role in the pathogenesis of human immunodeficiency virus (HIV)-associated nephropathy. A key question concerns the mechanism(s) by which the HIV-1 genome alters the phenotype of the highly specialized, terminally differentiated podocytes. Here, using an in vitro system of conditionally immortalized differentiated human podocytes (CIDHPs), we document a pivotal role for the p66ShcA protein in HIV-1-induced reactive oxygen species generation and CIDHP apoptosis. CIDHP transfected with truncated HIV-1 construct (NL4-3) exhibit increased reactive oxygen species metabolism, DNA strand breaks, and a 5-fold increase in apoptosis, whereas the opposite was true for NL4-3/CIDHP co-transfected with mu-36p66ShcA (micro-36) dominant negative expression vector or isoform-specific p66-small interfering RNA. Phosphorylation at Ser-36 of the wild type p66ShcA protein, required for p66ShcA redox function and inhibition of the potent stress response regulator Foxo3a, was unchanged in micro-36/NL4-3/CIDHP but increased in NL4-3/CIDHP. Acute knockdown of Foxo3a by small interfering RNA induced a 50% increase in micro-36/NL4-3/CIDHP apoptosis, indicating that Foxo3a-dependent responses promote the survival phenotype in micro-36 cells. We conclude that inhibition of p66ShcA redox activity prevents generation of HIV-1 stress signals and activation of the CIDHP apoptosis program. |
DOI | 10.1074/jbc.M109.008482 |
Alternate Journal | J Biol Chem |
PubMed ID | 19383602 |
PubMed Central ID | PMC2713565 |
Grant List | G0800200 / MRC_ / Medical Research Council / United Kingdom R01 DA012111 / DA / NIDA NIH HHS / United States R01 DK073793 / DK / NIDDK NIH HHS / United States R01 NS055644 / NS / NINDS NIH HHS / United States 1R01 DK073793 / DK / NIDDK NIH HHS / United States 1R01 HL072852 / HL / NHLBI NIH HHS / United States R01 HL072852 / HL / NHLBI NIH HHS / United States P01 NS043980-06A1 / NS / NINDS NIH HHS / United States P01 NS043980 / NS / NINDS NIH HHS / United States R01DA12111 / DA / NIDA NIH HHS / United States |
Related Faculty:
Surya V. Seshan, M.D.