- MeSH
- infekční lékařství * MeSH
- interakce hostitele a patogenu MeSH
- Publikační typ
- úvodní články MeSH
The cellular role of breast carcinoma-associated protein (BCA3), also known as A-kinase-interacting protein 1 (AKIP-1), is not fully understood. Recently, we reported that full-length, but not C-terminally truncated, BCA3 is incorporated into virions of Mason-Pfizer monkey virus, and that BCA3 enhances HIV-1 protease-induced apoptosis. In the present study, we report that BCA3 is associated with purified and subtilisin-treated HIV particles. Using a combination of immune-based methods and confocal microscopy, we show that the C-terminus of BCA3 is required for packaging into HIV-1 particles. However, we were unable to identify an HIV-1 binding domain for BCA3, and we did not observe any effect of incorporated BCA3 on HIV-1 infectivity. Interestingly, the BCA3 C-terminus was previously identified as a binding site for the catalytic subunit of protein kinase A (PKAc), a cellular protein that is specifically packaged into HIV-1 particles. Based on our analysis of PKAc⁻BCA3 interactions, we suggest that BCA3 incorporation into HIV-1 particles is mediated by its ability to interact with PKAc.
- MeSH
- adaptorové proteiny signální transdukční genetika metabolismus MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- HIV-1 metabolismus fyziologie MeSH
- jaderné proteiny genetika metabolismus MeSH
- lidé MeSH
- proteinkinasy závislé na cyklickém AMP metabolismus MeSH
- replikace viru genetika MeSH
- sestavení viru MeSH
- vazba proteinů MeSH
- virion metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Despite successful vaccination programs and effective treatments for some viral infections, humans are still losing the battle with viruses. Persisting human pandemics, emerging and re-emerging viruses, and evolution of drug-resistant strains impose continuous search for new antiviral drugs. A combination of detailed information about the molecular organization of viruses and progress in molecular biology and computer technologies has enabled rational antivirals design. Initial step in establishing efficacy of new antivirals is based on simple methods assessing inhibition of the intended target. We provide here an overview of biochemical and cell-based assays evaluating the activity of inhibitors of clinically important viruses.
- MeSH
- antivirové látky farmakologie MeSH
- fyziologie virů účinky léků MeSH
- inhibitory enzymů farmakologie MeSH
- interakce hostitele a patogenu účinky léků MeSH
- internalizace viru účinky léků MeSH
- kapsida účinky léků metabolismus MeSH
- lidé MeSH
- preklinické hodnocení léčiv metody MeSH
- replikace viru účinky léků MeSH
- rychlé screeningové testy metody MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Klíčová slova
- nezralé částice, inhibitory schopnosti tvořit částice,
- MeSH
- fluorescenční mikroskopie MeSH
- genové produkty gag - virus lidské imunodeficience MeSH
- HIV infekce epidemiologie etiologie terapie MeSH
- HIV-proteasa MeSH
- HIV genetika patogenita růst a vývoj účinky léků MeSH
- látky proti HIV * farmakologie MeSH
- léková rezistence MeSH
- lidé MeSH
- mutantní proteiny MeSH
- objevování léků * metody MeSH
- techniky in vitro MeSH
- testy rezistence k nukleáze MeSH
- vysoce aktivní antiretrovirová terapie ekonomika škodlivé účinky MeSH
- Check Tag
- lidé MeSH
We identified breast cancer-associated protein (BCA3) as a novel binding partner of Mason-Pfizer monkey virus (MPMV) protease (PR). The interaction was confirmed by co-immunoprecipitation and immunocolocalization of MPMV PR and BCA3. Full-length but not C-terminally truncated BCA3 was incorporated into MPMV virions. We ruled out the potential role of the G-patch domain, a glycine-rich domain located at the C terminus of MPMV PR, in BCA3 interaction and virion incorporation. Expression of BCA3 did not affect MPMV particle release and proteolytic processing; however, it slightly increased MPMV infectivity.
- MeSH
- adaptorové proteiny signální transdukční chemie genetika metabolismus MeSH
- druhová specificita MeSH
- endopeptidasy chemie genetika metabolismus MeSH
- HEK293 buňky MeSH
- interakční proteinové domény a motivy MeSH
- jaderné proteiny chemie genetika metabolismus MeSH
- lidé MeSH
- Masonův-Pfizerův opičí virus enzymologie genetika MeSH
- molekulární sekvence - údaje MeSH
- rekombinantní proteiny chemie genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- vazba proteinů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Apoptosis is one of the presumptive causes of CD4+ T cell depletion during HIV infection and progression to AIDS. However, the precise role of HIV-1 in this process remains unexplained. HIV-1 protease (PR) has been suggested as a possible factor, but a direct link between HIV-1 PR enzymatic activity and apoptosis has not been established. RESULTS: Here, we show that expression of active HIV-1 PR induces death in HeLa and HEK-293 cells via the mitochondrial apoptotic pathway. This conclusion is based on in vivo observations of the direct localization of HIV-1 PR in mitochondria, a key player in triggering apoptosis. Moreover, we observed an HIV-1 PR concentration-dependent decrease in mitochondrial membrane potential and the role of HIV-1 PR in activation of caspase 9, PARP cleavage and DNA fragmentation. In addition, in vitro data demonstrated that HIV-1 PR mediates cleavage of mitochondrial proteins Tom22, VDAC and ANT, leading to release of AIF and Hsp60 proteins. By using yeast two-hybrid screening, we also identified a new HIV-1 PR interaction partner, breast carcinoma-associated protein 3 (BCA3). We found that BCA3 accelerates p53 transcriptional activity on the bax promoter, thus elevating the cellular level of pro-apoptotic Bax protein. CONCLUSION: In summary, our results describe the involvement of HIV-1 PR in apoptosis, which is caused either by a direct effect of HIV-1 PR on mitochondrial membrane integrity or by its interaction with cellular protein BCA3.
- MeSH
- adaptorové proteiny signální transdukční genetika metabolismus MeSH
- apoptóza genetika MeSH
- buněčné linie MeSH
- CD4-pozitivní T-lymfocyty metabolismus MeSH
- fragmentace DNA MeSH
- HEK293 buňky MeSH
- HeLa buňky MeSH
- HIV infekce genetika metabolismus MeSH
- HIV-1 genetika metabolismus MeSH
- HIV-proteasa genetika metabolismus MeSH
- jaderné proteiny genetika metabolismus MeSH
- lidé MeSH
- mitochondriální proteiny genetika metabolismus MeSH
- mitochondrie genetika metabolismus MeSH
- nádorové buněčné linie MeSH
- nádorový supresorový protein p53 genetika metabolismus MeSH
- promotorové oblasti (genetika) genetika MeSH
- protein X asociovaný s bcl-2 genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
