- MeSH
- geny gag MeSH
- kur domácí MeSH
- ptačí sarkom * MeSH
- sekvence aminokyselin MeSH
- Viverridae * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- dopisy MeSH
- komentáře MeSH
During the last few decades, the treatment of HIV-infected patients by highly active antiretroviral therapy, including protease inhibitors (PIs), has become standard. Here, we present results of analysis of a patient-derived, multiresistant HIV-1 CRF02_AG recombinant strain with a highly mutated protease (PR) coding sequence, where up to 19 coding mutations have accumulated in the PR. The results of biochemical analysis in vitro showed that the patient-derived PR is highly resistant to most of the currently used PIs and that it also exhibits very poor catalytic activity. Determination of the crystal structure revealed prominent changes in the flap elbow region and S1/S1' active site subsites. While viral loads in the patient were found to be high, the insertion of the patient-derived PR into a HIV-1 subtype B backbone resulted in reduction of infectivity by 3 orders of magnitude. Fitness compensation was not achieved by elevated polymerase (Pol) expression, but the introduction of patient-derived gag and pol sequences in a CRF02_AG backbone rescued viral infectivity to near wild-type (wt) levels. The mutations that accumulated in the vicinity of the processing sites spanning the p2/NC, NC/p1, and p6pol/PR proteins lead to much more efficient hydrolysis of corresponding peptides by patient-derived PR in comparison to the wt enzyme. This indicates a very efficient coevolution of enzyme and substrate maintaining high viral loads in vivo under constant drug pressure.
- MeSH
- buněčné linie MeSH
- genové produkty gag - virus lidské imunodeficience genetika MeSH
- genové produkty pol - virus lidské imunodeficience genetika MeSH
- geny gag MeSH
- geny pol MeSH
- HEK293 buňky MeSH
- HIV infekce farmakoterapie virologie MeSH
- HIV-1 genetika izolace a purifikace fyziologie MeSH
- HIV-proteasa chemie genetika metabolismus MeSH
- inhibitory HIV-proteasy terapeutické užití MeSH
- krystalografie rentgenová MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- mutace MeSH
- peptidové fragmenty genetika MeSH
- virová léková rezistence genetika MeSH
- virová nálož MeSH
- vysoce aktivní antiretrovirová terapie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, Non-P.H.S. MeSH
- MeSH
- genetické testování MeSH
- geny gag MeSH
- lidé MeSH
- mutace MeSH
- olivopontocerebelární atrofie genetika patologie MeSH
- spinocerebelární ataxie epidemiologie genetika patologie MeSH
- Check Tag
- lidé MeSH
- Geografické názvy
- Česká republika MeSH
- MeSH
- endopeptidasy biosyntéza izolace a purifikace metabolismus MeSH
- gag-pol fúzové proteiny biosyntéza izolace a purifikace metabolismus MeSH
- geny gag MeSH
- geny pol MeSH
- virus ptačí myeloblastózy enzymologie genetika MeSH
- viry ptačího sarkomu enzymologie genetika MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- geny gag MeSH
- proteosyntéza MeSH
- RNA izolace a purifikace MeSH
- virus ptačí myeloblastózy genetika MeSH
- Publikační typ
- techniky in vitro MeSH
