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Molecular and Cellular Biochemistry

, Volume 345, Issue 1–2, pp 105–118 | Cite as

Effect of the redox state on HIV-1 tat protein multimerization and cell internalization and trafficking

  • Raffaella Pierleoni
  • Michele Menotta
  • Antonella Antonelli
  • Carla Sfara
  • Giordano Serafini
  • Sabrina Dominici
  • Maria Elena Laguardia
  • Annalisa Salis
  • Gianluca Damonte
  • Lucia Banci
  • Marco Porcu
  • Paolo Monini
  • Barbara Ensoli
  • Mauro MagnaniEmail author
Article

Abstract

The redox state of the cysteine-rich region of the HIV Tat protein is known to play a crucial role in Tat biological activity. In this article, we show that Tat displays two alternative functional states depending on the presence of either one or three reduced sulphydryl groups in the cysteine-rich region, respectively. Using different approaches, a disulfide pattern has been defined for the Tat protein and a specific DTT-dependent breaking order of disulfide bonds highlighted. The Tat redox state deeply influences macrophage protein uptake. Immunoistochemistry analysis shows that the oxidized protein does not enter cells, whereas partially reduced protein reaches the cytosol and, to a limited extent, the nucleus. Finally electrophoretic analysis shows Tat high-molecular weight multi-aggregation, resulting in the loss of biological activity. This is due to strong electrostatic and metal-binding interactions, whereas Tat dimerization involves metal-binding interactions as well as disulfide bond formation.

Keywords

HIV Tat Redox state Cell internalization 

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Copyright information

© Springer Science+Business Media, LLC. 2010

Authors and Affiliations

  • Raffaella Pierleoni
    • 1
  • Michele Menotta
    • 1
  • Antonella Antonelli
    • 1
  • Carla Sfara
    • 1
  • Giordano Serafini
    • 1
  • Sabrina Dominici
    • 1
  • Maria Elena Laguardia
    • 2
  • Annalisa Salis
    • 3
  • Gianluca Damonte
    • 3
  • Lucia Banci
    • 4
  • Marco Porcu
    • 4
  • Paolo Monini
    • 5
  • Barbara Ensoli
    • 5
  • Mauro Magnani
    • 1
    Email author
  1. 1.Department of Molecular SciencesUniversity of Urbino “Carlo Bo”UrbinoItaly
  2. 2.Diatheva srlFanoItaly
  3. 3.Department of Experimental Medicine and Center of Excellence for Biomedical ResearchUniversity of GenovaGenovaItaly
  4. 4.Centro Risonanze MagneticheUniversity of FlorenceSesto Fiorentino FlorenceItaly
  5. 5.Centro Nazionale AIDSIstituto Superiore di SanitàRomeItaly

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