It was proposed that Ato1p, Ato2p and Ato3p have a role in ammonia production by Saccharomyces cerevisiae colonies (Palkova et al., Mol Biol Cell 13: 3901-3914, 2002). In this study, we show that all three Ato proteins localise to the plasma membrane and their appearance correlates with the beginning of ammonia release. The expression of ATO genes is controlled by ammonia. All three Ato-GFP proteins associate with detergent-resistant membranes; two of them, Ato1p-GFP and Ato3p-GFP, localise to patches visible under the fluorescence microscope. In contrast with Ato3p-GFP which forms stable patches, the formation of those of Ato1p-GFP is pH dependent. Ato1p-GFP patches form at pH above 6 and they disappear at pH 5 or lower. Both changes, Ato1p-GFP clustering and patches spreading are reversible. The Ato1p-GFP spreading at low pH is independent on endocytosis. These data suggest that besides the ammonia induction of Ato protein synthesis, pH may rapidly regulate Ato1p function.

• MeSH
• amoniak metabolismus   MeSH
• buněčná membrána metabolismus   MeSH
• detergenty metabolismus   MeSH
• kompartmentace buňky * MeSH
• koncentrace vodíkových iontů MeSH
• kvartérní amoniové sloučeniny metabolismus   MeSH
• membránové proteiny metabolismus   MeSH
• membránové transportní proteiny metabolismus   MeSH
• rekombinantní fúzní proteiny metabolismus   MeSH
• Saccharomyces cerevisiae - proteiny metabolismus   MeSH
• Saccharomyces cerevisiae cytologie   metabolismus   MeSH
• transport proteinů MeSH
• zelené fluorescenční proteiny metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• amoniak MeSH
• ATO1 protein, S cerevisiae MeSH Prohlížeč
• ATO2 protein, S cerevisiae MeSH Prohlížeč
• ATO3 protein, S cerevisiae MeSH Prohlížeč
• detergenty MeSH
• kvartérní amoniové sloučeniny MeSH
• membránové proteiny MeSH
• membránové transportní proteiny MeSH
• rekombinantní fúzní proteiny MeSH
• Saccharomyces cerevisiae - proteiny MeSH
• zelené fluorescenční proteiny MeSH

Precise quantification and quality characterisation of isolated RNAs are prerequisites for their further exploitation in genome-wide microarrays, Northern blots, cDNA library preparation and others. Our data indicate that RNA analyses using Agilent RNA Nano Assay exhibit several advantages when compared with those performed on ethidium bromide-stained agarose gel electrophoresis or on a spectrophotometer. The RNA Nano Assay makes it possible to estimate RNA concentrations in the range from 1000 ng microl(-1) to 17 ng microl(-1). The presence of impurities including traces of DNA within RNA samples does not influence the concentration measurements. Like agarose gel electrophoresis, RNA Nano Assay allows to analyse RNAs dissolved in formamide and therefore protected against RNase action. Moreover, it allows a clearer distinction of partially degraded samples. The limitation of RNA Nano Assay is the impossibility to detect and to analyse double-stranded RNAs.

• MeSH
• elektroforéza v agarovém gelu metody   MeSH
• exprese genu genetika   MeSH
• fungální RNA analýza   MeSH
• polymerázová řetězová reakce přístrojové vybavení   metody   MeSH
• reagenční diagnostické soupravy * MeSH
• Saccharomyces cerevisiae genetika   MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• fungální RNA MeSH
• reagenční diagnostické soupravy * MeSH