Modulation of HIV-1 Gag NC/p1 cleavage efficiency affects protease inhibitor resistance and viral replicative capacity

Retrovirology

Monique Nijhuis³  Charles AB Boucher²  Dorien de Jong³  Pauline J Schipper³  Axel Fun³  Martin Lepšík¹  Dan Andersson¹  Noortje M van Maarseveen³ 

[1] Gilead Sciences and IOCB Research Center Prague, Institute of Organic Chemistry and Biochemistry, v.v.i., Academy of Science of the Czech Republic, Flemingovo n.2, 166 10 Praha 6, Czech Republic;Dept. of Virology, Erasmus Medical Center, Dr. Molewaterplein 50, 3015 GE Rotterdam, the Netherlands;Dept. of Medical Microbiology, Virology, University Medical Center Utrecht, Heidelberglaan 100 (HP G04.614), 3584 CX Utrecht, the Netherlands

关键词: NC/p1;    Replicative capacity;    Cleavage;    Gag;    Resistance;    Protease;    HIV-1;   

Others  :  1209316
DOI  :  10.1186/1742-4690-9-29

 received in 2011-10-21, accepted in 2012-04-01,  发布年份 2012

【 摘 要 】

Background

Mutations in the substrate of HIV-1 protease, especially changes in the NC/p1 cleavage site, can directly contribute to protease inhibitor (PI) resistance and also compensate for defects in viral replicative capacity (RC) due to a drug resistant protease. These NC/p1 changes are known to enhance processing of the Gag protein. To investigate the capacity of HIV-1 to modulate Gag cleavage and its consequences for PI resistance and RC, we performed a detailed enzymatic and virological analysis using a set of PI resistant NC/p1 variants (HXB2^431V, HXB2^436E+437T, HXB2^437T and HXB2^437V).

Results

Here, we demonstrate that single NC/p1 mutants, which displayed only a slight increase in PI resistance did not show an obvious change in RC. In contrast, the double NC/p1 mutant, which displayed a clear increase in processing efficiency and PI resistance, demonstrated a clear reduction in RC. Cleavage analysis showed that a tridecameric NC/p1 peptide representing the double NC/p1 mutant was cleaved in two specific ways instead of one.

The observed decrease in RC for the double NC/p1 mutant (HXB2^436E+437T) could (partially) be restored by either reversion of the 436E change or by acquisition of additional changes in the NC/p1 cleavage site at codon 435 or 438 as was revealed during in vitro evolution experiments. These changes not only restored RC but also reduced PI resistance levels. Furthermore these changes normalized Gag processing efficiency and obstructed the novel secondary cleavage site observed for the double NC/p1 mutant.

Conclusions

The results of this study clearly demonstrate that HIV-1 can modulate Gag processing and thereby PI resistance. Distinct increases in Gag cleavage and PI resistance result in a reduced RC that can only be restored by amino acid changes in NC/p1 which reduce Gag processing to an optimal rate.

【 授权许可】

   
2012 van Maarseveen et al; licensee BioMed Central Ltd.

【 图 表 】

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