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1.
A new planarian extrachromosomal virus-like element revealed by subtraction hybridization 总被引:1,自引:0,他引:1
A combination of suppression subtraction hybridization (SSH) and a new technique of mirror orientation selection (MOS) was used to compare the total DNA for two, sexual (SR) and asexual (AR), races of freshwater planarian Giradia tigrina. Several race-specific DNA fragments were found. A new element termed planarian extrachromosomal virus-like element (PEVE) was revealed in AR. The PEVE genome contains two unique regions, Ul and Us, which are flanked by inverted repeats. Two variants observed for the PEVE genome differ in combination of single- and double-stranded regions corresponding to Ul and Us. The PEVE genome codes for two helicases, one homologous to the circovirus replication initiation protein (Rep) and one corresponding to the helicase domain of papillomavirus E1. PEVE is nonuniformly distributed though the planarian body and is possibly replicated only in certain parenchymal cells. 相似文献
2.
Yanushevich Yu. G. Bulina M. E. Gurskaya N. G. Savitskii A. P. Lukyanov K. A. 《Russian Journal of Bioorganic Chemistry》2002,28(4):274-277
Site-directed mutagenesis was used to study the structural basis of color diversity of fluorescent proteins by the example of two closely related proteins from one organism (coral polyp Zoanthus sp.), one of which produces green and the other, yellow fluorescence. As a result, the following conversions of emission colors were performed: from yellow to green, from yellow to a dual color (yellow and green), and from green to yellow. The saltatory character of the spectral transitions and the manifestation of the dual-color fluorescence suggest that chemically different fluorophores are responsible for the green and yellow fluorescence. The simultaneous presence of three residues, Gly63, Lys65, and Asp68, is necessary for the efficient formation of the yellow rather than green fluorophore. 相似文献
3.
Bulina ME Lukyanov KA Yampolsky IV Chudakov DM Staroverov DB Shcheglov AS Gurskaya NG Lukyanov S 《The Journal of biological chemistry》2004,279(42):43367-43370
The nature of coloration in many marine animals remains poorly investigated. Here we studied the blue pigment of a scyfoid jellyfish Rhizostoma pulmo and determined it to be a soluble extracellular 30-kDa chromoprotein with a complex absorption spectrum peaking at 420, 588, and 624 nm. Furthermore, we cloned the corresponding cDNA and confirmed its identity by immunoblotting and mass spectrometry experiments. The chromoprotein, named rpulFKz1, consists of two domains, a Frizzled cysteine-rich domain and a Kringle domain, inserted into one another. Generally, Frizzleds are members of a basic Wnt signal transduction pathway investigated intensely with regard to development and cancerogenesis. Kringles are autonomous structural domains found throughout the blood clotting and fibrinolytic proteins. Neither Frizzled and Kringle domains association with any type of coloration nor Kringle intrusion into Frizzled sequence was ever observed. Thus, rpulFKz1 represents a new class of animal pigments, whose chromogenic group remains undetermined. The striking homology between a chromoprotein and members of the signal transduction pathway provides a novel node in the evolution track of growth factor-mediated morphogenesis compounds. 相似文献
4.
A genetically encoded photosensitizer 总被引:2,自引:0,他引:2
Bulina ME Chudakov DM Britanova OV Yanushevich YG Staroverov DB Chepurnykh TV Merzlyak EM Shkrob MA Lukyanov S Lukyanov KA 《Nature biotechnology》2006,24(1):95-99
Photosensitizers are chromophores that generate reactive oxygen species (ROS) upon light irradiation. They are used for inactivation of specific proteins by chromophore-assisted light inactivation (CALI) and for light-induced cell killing in photodynamic therapy. Here we report a genetically encoded photosensitizer, which we call KillerRed, developed from the hydrozoan chromoprotein anm2CP, a homolog of green fluorescent protein (GFP). KillerRed generates ROS upon irradiation with green light. Whereas known photosensitizers must be added to living systems exogenously, KillerRed is fully genetically encoded. We demonstrate the utility of KillerRed for light-induced killing of Escherichia coli and eukaryotic cells and for inactivating fusions to beta-galactosidase and phospholipase Cdelta1 pleckstrin homology domain. 相似文献
5.
Field SF Bulina MY Kelmanson IV Bielawski JP Matz MV 《Journal of molecular evolution》2006,62(3):332-339
Here we investigate the evolutionary scenarios that led to the appearance of fluorescent color diversity in reef-building
corals. We show that the mutations that have been responsible for the generation of new cyan and red phenotypes from the ancestral
green were fixed with the help of positive natural selection. This fact strongly suggests that the color diversity is a product
of adaptive evolution. An unexpected finding was a set of residues arranged as an intermolecular binding interface, which
was also identified as a target of positive selection but is nevertheless not related to color diversification. We hypothesize
that multicolored fluorescent proteins evolved as part of a mechanism regulating the relationships between the coral and its
algal endosymbionts (zooxanthellae). We envision that the effect of the proteins’ fluorescence on algal physiology may be
achieved not only through photosynthesis modulation, but also through regulatory photosensors analogous to phytochromes and
cryptochromes of higher plants. Such a regulation would require relatively subtle, but spectrally precise, modifications of
the light field. Evolution of such a mechanism would explain both the adaptive diversification of colors and the coevolutionary
chase at the putative algae-protein binding interface in coral fluorescent proteins.
Electronic Supplementary Material Electronic Supplementary material is available for this article at
and accessible for authorised users.
[Reviewing Editor: Dr. Rasmus Neilsen] 相似文献
6.
Ianushevich IuG Bulina ME Gurskaia NG Savitskiĭ AP Luk'ianov KA 《Bioorganicheskaia khimiia》2002,28(4):303-307
Site-directed mutagenesis was used to study the structural basis of color diversity of fluorescent proteins by the example of two closely related proteins from one organism (coral polyp Zoanthus sp.), one of which produces green and the other, yellow fluorescence. As a result, the following conversions of emission colors were performed: from yellow to green, from yellow to a dual color (yellow and green), and from green to yellow. The saltatory character of the spectral transitions and the manifestation of the dual-color fluorescence suggest that chemically different fluorophores are responsible for the green and yellow fluorescence. The simultaneous presence of three residues, Gly63, Lys65, and Asp68, is necessary for the efficient formation of the yellow rather than green fluorophore. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2002, vol. 28, no. 4; see also http://www.maik.ru. 相似文献
7.
Merzlyak EM Goedhart J Shcherbo D Bulina ME Shcheglov AS Fradkov AF Gaintzeva A Lukyanov KA Lukyanov S Gadella TW Chudakov DM 《Nature methods》2007,4(7):555-557
Fluorescent proteins have become extremely popular tools for in vivo imaging and especially for the study of localization, motility and interaction of proteins in living cells. Here we report TagRFP, a monomeric red fluorescent protein, which is characterized by high brightness, complete chromophore maturation, prolonged fluorescence lifetime and high pH-stability. These properties make TagRFP an excellent tag for protein localization studies and fluorescence resonance energy transfer (FRET) applications. 相似文献
8.
Bulina ME Lukyanov KA Britanova OV Onichtchouk D Lukyanov S Chudakov DM 《Nature protocols》2006,1(2):947-953
The phototoxic red fluorescent GFP-like protein KillerRed has recently been described. The phototoxicity of KillerRed exceeds that of EGFP by at least 1,000-fold, making it the first fully genetically encoded photosensitizer. KillerRed opens up new possibilities for precise light-induced cell killing and target protein inactivation. Because KillerRed is encoded by a gene, it can be expressed in a spatially and temporally regulated manner, under a chosen promoter, and fused with the desired protein of interest or localization signal. Here we provide a protocol for target protein inactivation in cell culture using KillerRed. As KillerRed is a new tool, the protocol focuses on aspects that will allow users to maximize the potential of this protein, guiding the design of chimeric constructs, recommended control experiments and preferred illumination parameters. The protocol, which describes target protein visualization and subsequent inactivation, is a 2- or 3-d procedure. 相似文献
9.
Denis V Rebrikov Maria E Bulina Ekaterina A Bogdanova Loura L Vagner Sergey A Lukyanov 《BMC genomics》2002,3(1):15
Background
Freshwater planarians are widely used as models for investigation of pattern formation and studies on genetic variation in populations. Despite extensive information on the biology and genetics of planaria, the occurrence and distribution of viruses in these animals remains an unexplored area of research. 相似文献10.
Oswald F Schmitt F Leutenegger A Ivanchenko S D'Angelo C Salih A Maslakova S Bulina M Schirmbeck R Nienhaus GU Matz MV Wiedenmann J 《The FEBS journal》2007,274(4):1102-1109
For a variety of coral species, we have studied the molecular origin of their coloration to assess the contributions of host and symbiont pigments. For the corals Catalaphyllia jardinei and an orange-emitting color morph of Lobophyllia hemprichii, the pigments belong to a particular class of green fluorescent protein-like proteins that change their color from green to red upon irradiation with approximately 400 nm light. The optical absorption and emission properties of these proteins were characterized in detail. Their spectra were found to be similar to those of phycoerythrin from cyanobacterial symbionts. To unambiguously determine the molecular origin of the coloration, we performed immunochemical studies using double diffusion in gel analysis on tissue extracts, including also a third coral species, Montastrea cavernosa, which allowed us to attribute the red fluorescent coloration to green-to-red photoconvertible fluorescent proteins. The red fluorescent proteins are localized mainly in the ectodermal tissue and contribute up to 7.0% of the total soluble cellular proteins in these species. Distinct spatial distributions of green and cyan fluorescent proteins were observed for the tissues of M. cavernosa. This observation may suggest that differently colored green fluorescent protein-like proteins have different, specific functions. In addition to green fluorescent protein-like proteins, the pigments of zooxanthellae have a strong effect on the visual appearance of the latter species. 相似文献