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Ornithine decarboxylase (ODC), a homodimeric enzyme with a rate-limiting function in polyamine biosynthesis, is subject to a feedback control involving its selective proteolysis. Targeting of ODC monomers to the proteasome is mediated by ODC antizyme (OAZ), the expression of which is induced by high levels of polyamines. Here, we report our analysis of the N-terminal degron in Saccharomyces cerevisiae ODC and the mechanism of its antizyme-dependent targeting. This ∼ 45-residue domain of ODC [termed ODC degradation signal (ODS)] is essential for degradation of ODC. Extensive mutagenesis indicated that it is not a specific sequence within ODS that is important but, rather, its unstructured nature. Consistent with this conclusion, ODS could be functionally replaced by an unrelated unstructured domain. We show that increasing the distance of ODS to the rest of the ODC protein reduced the dependence on Oaz1 for targeting, indicating that exposure of ODS is critical for its function. Disruption of ODC dimers by introducing interface mutations, in contrast, was insufficient for targeting. Binding of Oaz1 to ODC monomers is thus required to activate ODS. Fusion of ODS to the N terminus of Ura3 was sufficient to convert it into a ubiquitin-independent substrate of the proteasome. By contrast, ODS failed to destabilize maltose-binding protein or dihydrofolate reductase, indicating that this degron only operates in an appropriate structural context that enables rapid unfolding. 相似文献
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Mekler V Kortkhonjia E Mukhopadhyay J Knight J Revyakin A Kapanidis AN Niu W Ebright YW Levy R Ebright RH 《Cell》2002,108(5):599-614
We have used systematic fluorescence resonance energy transfer and distance-constrained docking to define the three-dimensional structures of bacterial RNA polymerase holoenzyme and the bacterial RNA polymerase-promoter open complex in solution. The structures provide a framework for understanding sigma(70)-(RNA polymerase core), sigma(70)-DNA, and sigma(70)-RNA interactions. The positions of sigma(70) regions 1.2, 2, 3, and 4 are similar in holoenzyme and open complex. In contrast, the position of sigma(70) region 1.1 differs dramatically in holoenzyme and open complex. In holoenzyme, region 1.1 is located within the active-center cleft, apparently serving as a "molecular mimic" of DNA, but, in open complex, region 1.1 is located outside the active center cleft. The approach described here should be applicable to the analysis of other nanometer-scale complexes. 相似文献
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Salome Kiparoidze Nino Karumidze Ekaterine Bakuradze Irina Modebadze Levan Rusishvili Dimitri Kordzaia Diana Dzidziguri 《Molecular & cellular biomechanics : MCB》2021,18(3):131-138
According to the literature, different mechanisms and kinetics proceeding of regenerative growth has been established using the basic models of liver regeneration (after resection or chemically induced). Hence, in order to determine general regularities of the adaptive-compensatory processes in various pathological conditions, the processes taking place in the cholestatic liver of adult white rats during the first four days after common bile duct ligation have been studied. It has been shown that in cholestatic liver, compensatory-adaptive processes take place with different kinetics compared to those after resection. In particular, in response to the increased functional load caused by destructive processes during cholestasis, the liver, at an early stage, responds by simple division of high ploidy (binuclear tetraploid) cells and further provides their quantitative increase. The difference between the processes taking place in cholestatic and resected liver is more expressed on the third and fourth day after common bile duct ligation. In particular, 4c cells are still highest in cholestatic liver, while all ploidy cells are present in equal numbers in the regenerated liver after resection. This fact of compensatory growth characteristic for reparative regeneration was not detected in cholestatic liver at the mentioned date. 相似文献
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Leila Kvachadze Nana Balarjishvili Tamila Meskhi Ekaterine Tevdoradze Natia Skhirtladze Tamila Pataridze Revaz Adamia Temur Topuria Elizabeth Kutter Christine Rohde Mzia Kutateladze 《Microbial biotechnology》2011,4(5):643-650
In recent decades the increase in antibiotic‐resistant bacterial strains has become a serious threat to the treatment of infectious diseases. Drug resistance of Staphylococcus aureus has become a major problem in hospitals of many countries, including developed ones. Today the interest in alternative remedies to antibiotics, including bacteriophage treatment, is gaining new ground. Here, we describe the staphylococcal bacteriophage Sb‐1 – a key component of therapeutic phage preparation that was successfully used against staphylococcal infections during many years in the Former Soviet Union. This phage still reveals a high spectrum of lytic activity in vitro against freshly isolated, genetically different clinical samples (including methicillin‐resistant S. aureus) obtained from the local hospitals, as well as the clinics from different geographical areas. The sequence analyses of phage genome showed absence of bacterial virulence genes. A case report describes a promising clinical response after phage application in patient with cystic fibrosis and indicates the efficacy of usage of Sb‐1 phage against various staphylococcal infections. 相似文献
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Upstream promoter sequences and alphaCTD mediate stable DNA wrapping within the RNA polymerase-promoter open complex
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Cellai S Mangiarotti L Vannini N Naryshkin N Kortkhonjia E Ebright RH Rivetti C 《EMBO reports》2007,8(3):271-278
We show that the extent of stable DNA wrapping by Escherichia coli RNA polymerase (RNAP) in the RNAP-promoter open complex depends on the sequence of the promoter and, in particular, on the sequence of the upstream region of the promoter. We further show that the extent of stable DNA wrapping depends on the presence of the RNAP alpha-subunit carboxy-terminal domain and on the presence and length of the RNAP alpha-subunit interdomain linker. Our results indicate that the extensive stable DNA wrapping observed previously in the RNAP-promoter open complex at the lambda P(R) promoter is not a general feature of RNAP-promoter open complexes. 相似文献
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