cis-/trans-regulation
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The main intention of this study was the investigation of impact of natural biologically active ligands of nuclear retinoid/retinoid X receptors (all-trans and 9-cis retinoic acid) on proteomic pattern in human estrogen receptor negative breast cancer cell line MDA-MB-231. For this purpose, proteomic strategies based on bottom-up method were applied. The total cell proteins were extracted utilizing a commercially Radio-Immunoprecipitation Assay (RIPA) buffer and separated on 2D sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D SDS-PAGE). The proteins were subsequently digested in-gel by trypsin and their characterization was achieved by MALDI-TOF/TOF. By employing PDQuest™ software, we identified more than 50 proteins affected by retinoic acid isomers. For more information, 9 proteins which are associated with tumor process were selected. We determined that derivatives of retinoic acid led to significantly reduced level of proteins belonging to metabolic pathway (e.g. glyceraldehyde-3-phosphate dehydrogenase or pyruvate kinase 2) or to other cellular processes as apoptosis, regulation of transcription process or epithelial-mesenchymal transition (e.g. annexins, nucleoside diphosphate kinase B, vimentin). On the other hand all-trans retinoic acid treatment indicates up-regulated effect for heterogeneous nuclear ribonucleoprotein A2/B1.
- MeSH
- apoptóza účinky léků MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- epitelo-mezenchymální tranzice účinky léků MeSH
- heterogenní jaderné ribonukleoproteiny skupiny A-B genetika metabolismus MeSH
- lidé MeSH
- ligandy MeSH
- nádorové buněčné linie MeSH
- nádory prsu genetika MeSH
- proteomika * MeSH
- protinádorové látky farmakologie MeSH
- regulace genové exprese u nádorů * MeSH
- retinoidní X receptory genetika metabolismus MeSH
- tretinoin farmakologie MeSH
- upregulace MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The aim of this study was to contribute to clarifying the role of 6S RNA in the development and control of antibiotic biosynthesis in Streptomyces coelicolor. Due to the low energetic cost of gene silencing via 6S RNA, it is an easy and rapid means of down-regulating the expression of specific genes in response to signals from changes in the environment. The expression of 6S RNA in S. coelicolor is not constitutive, and its accumulation is adapted to changes in nutritional conditions. The 6S RNA of S. coelicolor is capable of interacting with RNA polymerase β β' subunits and is a template for the transcription of short pRNAs. Deletion of the ssrS gene from S. coelicolor affects the growth rate and causes changes in the expression of several pathway-specific genes involved in actinorhodin biosynthesis. The complementation of the ΔssrS strain with ssrS gene restored the wild-type levels of growth and actinorhodin production. We conclude that 6S RNA contributes to the optimization of cellular adaptation and is an important factor involved in the regulation of growth and expression of key genes for the biosynthesis of actinorhodin.
- MeSH
- antibakteriální látky biosyntéza MeSH
- bakteriální RNA genetika metabolismus MeSH
- delece genu MeSH
- DNA řízené RNA-polymerasy metabolismus MeSH
- nekódující RNA genetika metabolismus MeSH
- regulace genové exprese u bakterií * MeSH
- Streptomyces coelicolor genetika růst a vývoj metabolismus MeSH
- testy genetické komplementace MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Yeasts tightly regulate their intracellular concentrations of alkali metal cations. In Saccharomyces cerevisiae, the Nha1 Na(+) /H(+) -antiporter and Ena1 Na(+) -ATPase, mediate the efflux of toxic sodium and surplus potassium. We report the characterization of Candida glabrata CgCnh1 and CgEna1 homologues. Their substrate specificity and transport properties were compared upon expression in S. cerevisiae, and their function characterized directly in C. glabrata. The CgCnh1 antiporter and the CgEna1 ATPase transport both potassium and sodium when expressed in S. cerevisiae. CgEna1p fully complements the lack of S. cerevisiae own Na(+) -ATPases but the activity of the CgCnh1 antiporter is lower than that of ScNha1p. Candida glabrata deletion mutants and analyses of their phenotypes revealed that though both transporters have a broad substrate specificity, their function in C. glabrata cells is not the same. Their differing physiological roles are also reflected in their regulation of expression, CgENA1 is highly upregulated by an increased osmotic pressure or sodium concentration, whereas CgCNH1 is expressed constitutively. The Cnh1 antiporter is involved in the regulation of potassium content and the Ena1 ATPase in sodium detoxification of C. glabrata cells. This situation differs from S. cerevisiae, where the Nha1 antiporter and Ena ATPases both participate together in Na(+) detoxification and in the regulation of K(+) homeostasis.
- MeSH
- Candida glabrata genetika metabolismus fyziologie MeSH
- delece genu MeSH
- draslík metabolismus MeSH
- fungální proteiny genetika metabolismus MeSH
- homeostáza MeSH
- kationty metabolismus MeSH
- Na(+)-H(+) antiport genetika metabolismus MeSH
- regulace genové exprese u hub MeSH
- Saccharomyces cerevisiae metabolismus MeSH
- sodík metabolismus MeSH
- substrátová specifita MeSH
- testy genetické komplementace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- srovnávací studie MeSH
Heat shock transcription factors (Hsfs) are involved in multiple aspects of stress response and plant growth. However, their role during male gametophyte development is largely unknown, although the generative phase is the most sensitive and critical period in the plant life cycle. Based on a wide screen of T-DNA mutant lines, we identified the atren1 mutation (restricted to nucleolus1) in early male gametophytic gene At1g77570, which has the closest homology to HSFA5 gene, the member of a heat shock transcription factor (HSF) gene family. The mutation causes multiple defects in male gametophyte development in both structure and function. Because the mutation disrupts an early acting (AtREN1) gene, these pollen phenotype abnormalities appear from bicellular pollen stage to pollen maturation. Moreover, the consequent progamic phase is compromised as well as documented by pollen germination defects and limited transmission via male gametophyte. In addition, atren1/- plants are defective in heat stress (HS) response and produce notably higher proportion of aberrant pollen grains. AtREN1 protein is targeted specifically to the nucleolus that, together with the increased size of the nucleolus in atren1 pollen, suggests that it is likely to be involved in ribosomal RNA biogenesis or other nucleolar functions.
- MeSH
- alely MeSH
- Arabidopsis cytologie růst a vývoj metabolismus MeSH
- buněčné jadérko metabolismus MeSH
- DNA bakterií genetika MeSH
- DNA vazebné proteiny genetika metabolismus MeSH
- exony genetika MeSH
- fenotyp MeSH
- klíčení MeSH
- mutace genetika MeSH
- penetrance MeSH
- proteiny huseníčku genetika metabolismus MeSH
- pyl cytologie genetika růst a vývoj MeSH
- pylová láčka cytologie genetika růst a vývoj MeSH
- reakce na tepelný šok * genetika MeSH
- regulace genové exprese u rostlin MeSH
- segregace chromozomů genetika MeSH
- testy genetické komplementace MeSH
- transport proteinů MeSH
- vývojová regulace genové exprese MeSH
- zelené fluorescenční proteiny metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The metabolism of steroids and retinoids has been studied in detail for a long time, as these compounds are involved in a broad spectrum of physiological processes. Many enzymes participating in the conversion of such compounds are members of the short-chain dehydrogenase/reductase (SDR) superfamily. Despite great effort, there still remain a number of poorly characterized SDR proteins. According to various bioinformatics predictions, many of these proteins may play a role in the metabolism of steroids and retinoids. Dehydrogenase/reductase (SDR family) member 7 (DHRS7) is one such protein. In a previous study, we determined DHRS7 to be an integral membrane protein of the endoplasmic reticulum facing the lumen which has shown at least in vitro NADPH-dependent reducing activity toward several eobiotics and xenobiotics bearing a carbonyl moiety. In the present paper pure DHRS7 was used for a more detailed study of both substrate screening and an analysis of kinetics parameters of the physiologically important substrates androstene-3,17-dione, cortisone and all-trans-retinal. Expression patterns of DHRS7 at the mRNA as well as protein level were determined in a panel of various human tissue samples, a procedure that has enabled the first estimation of the possible biological function of this enzyme. DHRS7 is expressed in tissues such as prostate, adrenal glands, liver or intestine, where its activity could be well exploited. Preliminary indications show that DHRS7 exhibits dual substrate specificity recognizing not only steroids but also retinoids as potential substrates and could be important in the metabolism of these signalling molecules.
- MeSH
- androstendion metabolismus MeSH
- cirkulární dichroismus MeSH
- fylogeneze MeSH
- kinetika MeSH
- kortison metabolismus MeSH
- lidé MeSH
- oxidoreduktasy chemie genetika metabolismus MeSH
- regulace genové exprese enzymů MeSH
- retinaldehyd metabolismus MeSH
- steroidy metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The proper timing of flowering is essential for the adaptation of plant species to their ever-changing environments. The central position in a complex regulatory network is occupied by the protein FT, which acts as a florigen. We found that light, following a permissive period of darkness, was essential to induce the floral promoter CrFTL1 and to initiate flowering in seedlings of the short-day plant Chenopodium rubrum L. We also identified two novel CONSTANS-like genes in C. rubrum and observed their rhythmic diurnal and circadian expressions. Strong rhythmicity of expression suggested that the two genes might have been involved in the regulation of photoperiod-dependent processes, despite their inability to complement co mutation in A. thaliana. The CrCOL1 and CrCOL2 genes were downregulated by dark-light transition, regardless of the length of a preceding dark period. The same treatment activated the floral promoter CrFTL1. Light therefore affected CrCOL and CrFTL1 in an opposite manner. Both CrCOL genes and CrFTL1 displayed expression patterns unique among short-day plants. Chenopodium rubrum, the subject of classical physiological studies in the past, is emerging as a useful model for the investigation of flowering at the molecular level.
- MeSH
- Arabidopsis MeSH
- Chenopodium genetika růst a vývoj fyziologie MeSH
- florigen metabolismus MeSH
- fotoperioda MeSH
- květy růst a vývoj MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- promotorové oblasti (genetika) MeSH
- regulace genové exprese u rostlin * MeSH
- rostlinné proteiny genetika metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvenční seřazení MeSH
- semenáček růst a vývoj MeSH
- testy genetické komplementace MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Clostridium perfringens forms biofilms and spores that are a source of food contamination. In this study, the antibacterial activities of Lactobacillus plantarum culture supernatants (LP-S), LP-S fractions, and the plant-derived compound epigallocatechin gallate (EG) were evaluated. Specifically, their effects on the viability and biofilm-forming ability of C. perfringens were assessed. Moreover, the expression of quorum sensing-regulated genes associated with the pathogenesis of this microorganism and that of genes involved in biofilm formation was also investigated. The results showed that both EG and the LP-S exerted bactericidal activity against all C. perfringens strains tested. The minimal bactericidal concentration (MBC) of EG was 75 µg/mL for all strains but ranged from 61 to 121 µg of total protein per mL for LP-S. EG exerted only minor effects on biofilm formation, whereas LP-S, particularly its 10 and 30 K fractions, significantly reduced the biofilm-forming ability of all the strains. The antibiofilm activity of LP-S was lost following preincubation with proteases, suggesting that it was mediated by a proteinaceous molecule. The treatment of C. perfringens with either EG or LP-S did not change the transcript levels of two CpAL (C. perfringens quorum-sensing Agr-like system)-related genes, agrB and agrD, which are known to be involved in the regulation of biofilms, suggesting that LP-S exerted its biofilm inhibitory activity downstream of CpAL signaling. In summary, we demonstrated the bactericidal activity of EG and LP-S against C. perfringens and antibiofilm activity of LP-S at a subinhibitory dose. Our results suggested that these compounds can be further explored for food safety applications to control agents such as C. perfringens.
Metal accumulation in seeds is a prerequisite for germination and establishment of plants but also for micronutrient delivery to humans. To investigate metal transport processes and their interactions in seeds, we focused on METAL TOLERANCE PROTEIN8 (MTP8), a tonoplast transporter of the manganese (Mn) subclade of cation diffusion facilitators, which in Arabidopsis (Arabidopsis thaliana) is expressed in embryos of seeds. The x-ray fluorescence imaging showed that expression of MTP8 was responsible for Mn localization in subepidermal cells on the abaxial side of the cotyledons and in cortical cells of the hypocotyl. Accordingly, under low Mn availability, MTP8 increased seed stores of Mn, required for efficient seed germination. In mutant embryos lacking expression ofVACUOLAR IRON TRANSPORTER1(VIT1), MTP8 built up iron (Fe) hotspots inMTP8-expressing cells types, suggesting that MTP8 transports Fe in addition to Mn. Inmtp8 vit1double mutant seeds, Mn and Fe were distributed in all cell types of the embryo. An Fe transport function of MTP8 was confirmed by its ability to complement Fe hypersensitivity of a yeast mutant defective in vacuolar Fe transport. Imbibingmtp8-1mutant seeds in the presence of Mn or subjecting seeds to wet-dry cycles showed that MTP8 conferred Mn tolerance. During germination, MTP8 promoted reallocation of Fe from the vasculature. These results indicate that cell type-specific accumulation of Mn and Fe in seeds depends on MTP8 and that this transporter plays an important role in the generation of seed metal stores as well as for metal homeostasis and germination efficiency under challenging environmental conditions.
- MeSH
- Arabidopsis embryologie genetika metabolismus MeSH
- biologické modely MeSH
- genový knockout MeSH
- homeostáza * MeSH
- klíčení * genetika MeSH
- mangan metabolismus MeSH
- mutace genetika MeSH
- promotorové oblasti (genetika) genetika MeSH
- proteiny huseníčku metabolismus MeSH
- proteiny přenášející kationty metabolismus MeSH
- regulace genové exprese u rostlin MeSH
- Saccharomyces cerevisiae metabolismus MeSH
- semena rostlinná embryologie genetika MeSH
- spektrometrie rentgenová emisní MeSH
- testy genetické komplementace MeSH
- vývojová regulace genové exprese MeSH
- železo metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
