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Consensus HIV-1 subtype A integrase and its raltegravir-resistant variants: Design and characterization of the enzymatic properties.

In English

Författare

  • Olga Shadrina
  • Olga Krotova
  • Julia Agapkina
  • Ekaterina Knyazhanskaya
  • Sergey Korolev
  • Elizaveta Starodubova
  • Alecia Viklund
  • Vladimir Lukashov
  • Mauro Magnani
  • Patrik Medstrand
  • Vadim Karpov
  • Marina Gottikh
  • Maria Isaguliants
Visa allt

Summary, in English

Model studies of the subtype B and non-subtype B integrases are still required to compare their susceptibility to antiretroviral drugs, evaluate the significance of resistance mutations and identify the impact of natural polymorphisms on the level of enzymatic reactivity. We have therefore designed the consensus integrase of the HIV-1 subtype A strain circulating in the former Soviet Union territory (FSU-A) and two of its variants with mutations of resistance to the strand transfer inhibitor raltegravir. Their genes were synthesized, and expressed in E coli; corresponding His-tagged proteins were purified using the affinity chromatography. The enzymatic properties of the consensus integrases and their sensitivity to raltegravir were examined in a series of standard in vitro reactions and compared to the properties of the integrase of HIV-1 subtype B strain HXB2. The consensus enzyme demonstrated similar DNA-binding properties, but was significantly more active than HXB-2 integrase in the reactions of DNA cleavage and integration. All integrases were equally susceptible to inhibition by raltegravir and elvitegravir, indicating that the sporadic polymorphisms inherent to the HXB-2 enzyme have little effect on its susceptibility to drugs. Insensitivity of the mutated enzymes to the inhibitors of strand transfer occurred at a cost of a 30-90% loss of the efficacies of both 3'-processing and strand transfer. This is the first study to describe the enzymatic properties of the consensus integrase of HIV-1 clade A and the effects of the resistance mutations when the complex actions of sporadic sequence polymorphisms are excluded.

Publiceringsår

2014

Språk

Engelska

Sidor

92-101

Publikation/Tidskrift/Serie

Biochimie

Volym

102C

Issue

Mar 2

Dokumenttyp

Artikel i tidskrift

Förlag

Elsevier

Ämne

  • Biochemistry and Molecular Biology

Status

Published

Forskningsgrupp

  • Clinical Virology, Malmö

ISBN/ISSN/Övrigt

  • ISSN: 1638-6183