The mitochondrial rhomboid protease PARL regulates mitophagy by balancing intramembrane proteolysis of PINK1 and PGAM5. It has been implicated in the pathogenesis of Parkinson's disease, but its investigation as a possible therapeutic target is challenging in this context because genetic deficiency of PARL may result in compensatory mechanisms. To address this problem, we undertook a hitherto unavailable chemical biology strategy. We developed potent PARL-targeting ketoamide inhibitors and investigated the effects of acute PARL suppression on the processing status of PINK1 intermediates and on Parkin activation. This approach revealed that PARL inhibition leads to a robust activation of the PINK1/Parkin pathway without major secondary effects on mitochondrial properties, which demonstrates that the pharmacological blockage of PARL to boost PINK1/Parkin-dependent mitophagy is a feasible approach to examine novel therapeutic strategies for Parkinson's disease. More generally, this study showcases the power of ketoamide inhibitors for cell biological studies of rhomboid proteases.
- MeSH
- endopeptidasy MeSH
- lidé MeSH
- metaloproteasy genetika metabolismus MeSH
- mitochondriální proteiny metabolismus MeSH
- mitofagie MeSH
- Parkinsonova nemoc * farmakoterapie MeSH
- proteasy * MeSH
- proteinkinasy metabolismus MeSH
- ubikvitinligasy metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The membrane-embedded FtsH proteases found in bacteria, chloroplasts, and mitochondria are involved in diverse cellular processes including protein quality control and regulation. The genome of the model cyanobacterium Synechocystis sp PCC 6803 encodes four FtsH homologs designated FtsH1 to FtsH4. The FtsH3 homolog is present in two hetero-oligomeric complexes: FtsH2/3, which is responsible for photosystem II quality control, and the essential FtsH1/3 complex, which helps maintain Fe homeostasis by regulating the level of the transcription factor Fur. To gain a more comprehensive insight into the physiological roles of FtsH hetero-complexes, we performed genome-wide expression profiling and global proteomic analyses of Synechocystis mutants conditionally depleted of FtsH3 or FtsH1 grown under various nutrient conditions. We show that the lack of FtsH1/3 leads to a drastic reduction in the transcriptional response to nutrient stress of not only Fur but also the Pho, NdhR, and NtcA regulons. In addition, this effect is accompanied by the accumulation of the respective transcription factors. Thus, the FtsH1/3 complex is of critical importance for acclimation to iron, phosphate, carbon, and nitrogen starvation in Synechocystis.plantcell;31/12/2912/FX1F1fx1.
- MeSH
- aklimatizace genetika MeSH
- bakteriální proteiny genetika metabolismus MeSH
- dusík nedostatek metabolismus MeSH
- exprese genu MeSH
- fosfáty nedostatek metabolismus MeSH
- fosforylace MeSH
- fotosystém II - proteinový komplex chemie genetika metabolismus MeSH
- metaloproteasy genetika metabolismus MeSH
- mutace MeSH
- proteiny vázající fosfáty genetika metabolismus MeSH
- proteolýza MeSH
- proteom genetika metabolismus MeSH
- proteomika MeSH
- regulace genové exprese u bakterií genetika MeSH
- regulon genetika MeSH
- represorové proteiny genetika metabolismus MeSH
- ribozomální proteiny genetika metabolismus MeSH
- Synechocystis enzymologie metabolismus MeSH
- transkripční faktory genetika metabolismus MeSH
- uhlík nedostatek metabolismus MeSH
- živiny nedostatek metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
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- MeSH
- aspartátové proteasy genetika sekrece izolace a purifikace MeSH
- debridement * metody MeSH
- Diptera MeSH
- genová knihovna MeSH
- helmintoterapie MeSH
- hojení ran MeSH
- hybridizace in situ MeSH
- komplementární DNA genetika MeSH
- larva * enzymologie genetika růst a vývoj MeSH
- lidé MeSH
- metaloproteasy genetika izolace a purifikace sekrece MeSH
- nekróza terapie MeSH
- proteasy * genetika izolace a purifikace sekrece MeSH
- sekvenční analýza DNA MeSH
- serinové proteasy genetika sekrece izolace a purifikace MeSH
- slinné žlázy enzymologie MeSH
- trávicí systém enzymologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- práce podpořená grantem MeSH
Hepatic stellate cells (HSC) and liver myofibroblasts (MFB) are two cell populations most likely responsible for the synthesis of most connective tissue components in fibrotic liver. They differ in their origin and location, and possibly in patterns of gene expression. Normal and carbon tetrachloride-cirrhotic livers from rats were used to isolate HSC. Liver was perfused with pronase and collagenase solutions, followed by centrifugation of the cell suspension on a density gradient. HSC were quiescent 2 days after plating on plastic but they became activated after another 5 days in culture. When the culture was passaged 5 times, its character changed profoundly as HSC were replaced by MFB. Microarray analysis was used to determine gene expression in quiescent HSC, activated HSC and MFB. The expression of 49 genes coding for connective tissue proteins, proteoglycans, metalloproteinases and their inhibitors, growth factors and cellular markers was determined. The pattern of gene expression changed during HSC activation and there were distinct differences between HSC and MFB. Little difference between normal cells and cells isolated from cirrhotic liver was found.
- MeSH
- experimentální cirhóza jater metabolismus MeSH
- exprese genu MeSH
- extracelulární matrix chemie MeSH
- fibroblasty cytologie metabolismus účinky léků MeSH
- financování organizované MeSH
- imunohistochemie MeSH
- játra cytologie metabolismus účinky léků MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- messenger RNA biosyntéza účinky léků MeSH
- metaloproteasy genetika metabolismus MeSH
- myocyty hladké svaloviny cytologie metabolismus účinky léků MeSH
- otrava chloridem uhličitým MeSH
- pojivová tkáň metabolismus účinky léků MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- proteoglykany genetika metabolismus MeSH
- sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
- tkáňové inhibitory metaloproteinas MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
- MeSH
- apoptóza genetika účinky léků MeSH
- finanční podpora výzkumu jako téma MeSH
- hypoxie buňky fyziologie MeSH
- kardiomegalie patofyziologie MeSH
- metaloproteasy genetika metabolismus MeSH
- myokard enzymologie metabolismus patologie MeSH
- potkani Wistar anatomie a histologie MeSH
- synthasa oxidu dusnatého biosyntéza MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- srovnávací studie MeSH
