Interaction of HIV-1 Tat protein with heparin. Role of the backbone structure, sulfation, and size

Marco Rusnati, Daniela Coltrini, Pasqua Oreste, Giorgio Zoppetti, Adriana Albini, Douglas Noonan, Fabrizio D'Adda Di Fagagna, Mauro Giacca, Marco Presta

Research output: Contribution to journalArticle

165 Citations (Scopus)

Abstract

Human immunodeficiency virus type 1 (HIV-1) Tat protein is released from infected cells. Extracellular Tat enters the cell where it stimulates the transcriptional activity of HIV-long terminal repeat (LTR) and of endogenous genes. Heparin modulates the angiogenic (Albini, A., Benelli, R., Presta, M., Rusnati, M., Ziche, M., Rubartelli, A., Paglialunga, G., Bussolino, F., and Noonan, D. (1996) Oncogene 12, 289-297) and transcriptional (Mann, D. A., and Frankel, A.D. (1991) EMBO J. 10, 1733-1739) activity of extracellular Tat. Here we demonstrate that heparin binds specifically to recombinant HIV-1 Tat produced as glutathione S-transferase (GST) fusion protein and immobilized on glutathione-agarose beads. Heparin and heparan sulfate (HS), but not dermatan sulfate, chondroitin sulfates A and C, hyaluronic acid, and K5 polysaccharide, competed with 3H-labeled heparin for binding to immobilized GST-Tat and inhibited HIV-LTR transactivation induced by extracellular GST- Tat. Selective 2-O-, 6-O-, total-O-desulfation, or N-desulfation/N- acetylation dramatically reduced the capacity of heparin to bind GST-Tat. Totally-O-desulfated and 2-O-desulfated heparins also showed a reduced capacity to inhibit the transactivating activity of GST-Tat. Very low molecular weight heparins showed a significant decrease in their capacity to bind GST-Tat and to inhibit its LTR transactivating activity when compared with conventional 13.6-kDa heparin. However, when 3.0-kDa heparin was affinity chromatographed on immobilized GST-Tat to isolate binding and non- binding subfractions, the Tat-bound fraction was ≤1,000 times more potent than the unbound fraction in inhibiting the transactivating activity of GST- Tat. The results demonstrate that Tat interacts in a size-dependent manner with heparin/HS and that high affinity Tat-heparin interaction requires at least some 2-O-, 6-O-, and N-positions to be sulfated. The Tat binding activity of the glycosaminoglycans tested correlates with their capacity to affect the transactivating activity of extracellular Tat, indicating the possibility to design specific heparin/HS-like structures with Tat-antagonist activity.

Original languageEnglish
Pages (from-to)11313-11320
Number of pages8
JournalJournal of Biological Chemistry
Volume272
Issue number17
DOIs
Publication statusPublished - Apr 25 1997

Fingerprint

tat Gene Products
Viruses
Heparin
HIV-1
Glutathione Transferase
Heparitin Sulfate
HIV Long Terminal Repeat
Chondroitin Sulfates
Immobilized Proteins
Acetylation
Terminal Repeat Sequences
Low Molecular Weight Heparin
Hyaluronic Acid
Glycosaminoglycans
Oncogenes
Sepharose
Transcriptional Activation
Glutathione
Fusion reactions
Genes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Interaction of HIV-1 Tat protein with heparin. Role of the backbone structure, sulfation, and size. / Rusnati, Marco; Coltrini, Daniela; Oreste, Pasqua; Zoppetti, Giorgio; Albini, Adriana; Noonan, Douglas; D'Adda Di Fagagna, Fabrizio; Giacca, Mauro; Presta, Marco.

In: Journal of Biological Chemistry, Vol. 272, No. 17, 25.04.1997, p. 11313-11320.

Research output: Contribution to journalArticle

Rusnati, M, Coltrini, D, Oreste, P, Zoppetti, G, Albini, A, Noonan, D, D'Adda Di Fagagna, F, Giacca, M & Presta, M 1997, 'Interaction of HIV-1 Tat protein with heparin. Role of the backbone structure, sulfation, and size', Journal of Biological Chemistry, vol. 272, no. 17, pp. 11313-11320. https://doi.org/10.1074/jbc.272.17.11313
Rusnati, Marco ; Coltrini, Daniela ; Oreste, Pasqua ; Zoppetti, Giorgio ; Albini, Adriana ; Noonan, Douglas ; D'Adda Di Fagagna, Fabrizio ; Giacca, Mauro ; Presta, Marco. / Interaction of HIV-1 Tat protein with heparin. Role of the backbone structure, sulfation, and size. In: Journal of Biological Chemistry. 1997 ; Vol. 272, No. 17. pp. 11313-11320.
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AU - Albini, Adriana

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AU - D'Adda Di Fagagna, Fabrizio

AU - Giacca, Mauro

AU - Presta, Marco

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