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11.
Selenocysteine's mechanism of incorporation and evolution revealed in cDNAs of three glutathione peroxidases 总被引:5,自引:0,他引:5
G T Mullenbach A Tabrizi B D Irvine G I Bell J A Tainer R A Hallewell 《Protein engineering》1988,2(3):239-246
The nonsense codon, UGA, has for the first time recently been shown to encode selenocysteine in two proteins, mouse glutathione peroxidase (GSH-Px) (EC 1.11.1.9) and bacterial formate dehydrogenase. A co-translational rather than post-translational selenium-incorporation mechanism has been implicated. Furthermore, high expression levels of GSH-Px have suggested that suppression of termination is efficient and specific. We have isolated and characterized pituitary, kidney and placenta cDNAs for bovine, human and mouse GSH-Px respectively. It is demonstrated that this novel suppression event occurs in diverse tissues, in at least three mammalian species and at the translational step. Surprisingly, GSH-Px is shown to be extramitochondrially encoded, indicating a cytosolic suppression event rather than one utilizing the mitochondria's well-documented extended codon-reading ability. Sequence analysis reveals that a simple proximal contextual pattern responsible for readthrough does not exist. Analysis of predicted secondary structures of mRNAs, however, has revealed a conformation which may be unique to selenocysteine proteins and may prove useful as a tool for artificial incorporation of selenocysteines. A human intron for GSH-Px from an unspliced mRNA has been isolated whose position indicates an ancient, divergent evolutionary relationship with thioredoxin-S2, rather than an independent convergent one. 相似文献
12.
13.
14.
Jeanine Brederoo Piet de Wildt Corrie Popp-Snijders Robin F. Irvine Alan Musgrave Herman van den Ende 《Planta》1991,184(2):175-181
InChlamydomonas eugametos gametes, phosphatidylinositol 4-phosphate (PtdInsP) and phosphatidylinositol 4,5-bisphosphate (PtdInsP2) comprised 0.4 and 0.3% of the whole-cell phospholipids. They were concentrated in the plasma membrane around the cell body and were present in low concentrations in the flagellar membrane. When gametes were fed32PO
4
-
, the label was rapidly incorporated into PtdInsP and PtdInsP2 and only slowly incorporated into structural lipids such as phosphatidylethanolamine and phosphatidylglycerol. Similarly, when a pulse of32PO
4
-
was chased with PO
4
-
, the label was rapidly lost from the polyphosphoinositol lipids but not from the structural lipids. The major fatty acids in the polyphosphoinositides were C-22 carbon polyenoic acids (70%). The significance of these results in relationship to intracellular signalling via inositol phosphates and Ca2+ is discussed.Abbreviations InsP3
inositol 1,4,5-trisphosphate
- mt–/mt+
mating-type plus or minus
- PtdA
phosphatidic acid
- PtdEtn
phosphatidylethanolamine
- PtdGro
phosphatidylglycerol
- PtdIns
phosphatidylinositol
- PtdInsP
phosphatidylinositol 4-phosphate;
- PtdInsP2
phosphatidylinositol 4,5-bisphosphate
- TCA
trichloroacetic acid
We thank Frank Schuring for Fig. 5A and Susan Kenter, Hans Kruisselbrink, Saskia Bijvank and Nelleke Corbett for their enthousiastic assistance. 相似文献
15.
J. E. Irvine G. T. A. Benda B. L. Legendre G. R. Machado Jr 《Plant Cell, Tissue and Organ Culture》1991,26(2):115-125
This study investigates the frequency of apparent and permanent expression of marker change following two types of tissue culture, conventional callus and direct regeneration cultures, and for two markers it relates this frequency to that following breeding. Each clone was used for only one marker. After conventional callus culture, plants of the sugarcane clone Arundoid B, a clone having a growth habit with shortened internodes and leaves, were freed of this marker at a rate of 1 in 172 plants. Marker remission in a second clone with a leaf blotch was enhanced in the presence of a mutagen. Callus culture alone gave a remission rate of 1/280 plants, while treatment of callus with ethyl methanesulfonate gave a remission rate of 1/42 plants. Of two markers subjected to vegetative and sexual transmission, the first, a leaf marker, was stable in callus culture with no remissions; crossing with non-marker parents produced progeny with 54% lacking the marker. The second, a stalk marker (multibud), showed epigenetic effects during two generations of vegetative propagation; plants lacking the multibud marker produced vegetative progeny in which the marker reappeared. Nine crosses to nonmarker parents produced progeny of which an average of 29% had the marker. The use of stalk chimeras as markers demonstrated that passage through conventional callus or direct regeneration culture resulted in the loss of the donor phenotype in all plants regenerated. Phenotypic variation in plants derived from callus culture appears to arise from several sources; chimeral segregants, epigenetic transients, and mutational variants. 相似文献
16.
John H. Livesey Alan Carne Clifford H. G. Irvine Jane Ellis Margaret J. Evans Roger Smith Richard A. Donald 《Peptides》1991,12(6):1437-1440
A 41 amino acid peptide, probably identical in structure to human corticotropin releasing factor, was isolated from 70 equine hypothalami by methanol extraction, immunoaffinity chromatography and single step of reverse phase HPLC. The amino acid sequence was determined by gas phase sequence analysis. Probable carboxyl terminal amidation was demonstrated by similar retention times for equine and human corticotropin releasing factor on reverse phase HPLC at pH 8. The likely structure of equine corticotropin releasing factor is: Ser-Glu-Glu-Pro-Pro- Ile-Ser-Leu-Asp-Leu-Thr-Phe-His-Leu-Leu-Arg-Glu-Val-Leu-Glu-Met-Ala-Arg-Ala-Glu-Gln-Leu-Ala-Gln-Gln-Ala-His-Ser-Asn- Arg-Lys-Leu-Met-Glu-Ile-Ile-NH2. The purified peptide is equipotent with human corticotropin releasing factor in an in vitro bioassay and in a human plasma binding protein assay. 相似文献
17.
18.
Summary The general and ultrastructural organization of the heart of the elasmobranch, Scyllium stellare, was studied in normal and in anoxic animals. The rich coronary supply was revealed three-dimensionally by the use of corrosion casts, showing a thebesian system of coronary arterioles and capillaries in the thin, outer compact layer as well as in the predominant, inner spongy layer of trabeculae.Only the sinus venosus received a neuronal input of large bundles of granule-containing axons terminating at fenestrated regions of the endocardium and suggesting a neurohormonal function.A simple, tubular sarcoplasmic reticulum with flattened junctional cisternae was present in myocardial cells of 1–5 m diameter, which contained one or two bundles of myofibrils. The latter were closely apposed to the inner aspect of the plasmalemma. Mitochondria were located centrally in the cells, which were joined by unfolded desmosomes involving Z-band material.Long periods of anoxia were tolerated without loss of heart function, but at the expense of cytoplasmic glycogen. Lipid granules were abundant in all layers and chambers, notably in animals prepared in the summer. The lipid granules displayed a marked increase in electron density when the heart was incubated in a buffered oxalate solution prior to fixation. A glycogen-sparing effect of the lipids during anoxia was observed. 相似文献
19.
Karen B. Helle 《BBA》1971
1. Chromogranin A has been isolated from the washed membrane fraction of highly purified chromaffin granules after solubilization in Triton X-100. A hydrated molecular weight of 2.9·105 has been obtained for chromogranin AM which in its electrophoretic mobility and molecular volume appears very similar to chromogranin AI of the water-soluble fraction. 相似文献
20.
The susceptibility to Tedion of haploid and diploid-haploid mixtures of eggs of Tetranychus urticae Koch was examined. It was concluded for a normal susceptible strain that haploid eggs are more susceptible to Tedion than diploid eggs. This difference in tolerance between haploid and diploid eggs could not be established for a strain resistant to Tedion.Mass crosses between the susceptible and the resistant strain were made. Susceptible females, mated by resistant males, produce susceptible haploid and resistant diploid offspring. Resistant females, mated by susceptible males, gave a resistant offspring. Both sexes can also transmit resistance to Tedion. As there was a difference in tolerance between diploid offspring in the reciprocal crosses, it is assumed that either a maternal or a cytoplasmic component is also present in the genetical mechanism of Tedion-resistance.
Zusammenfassung Es wurde die Empfindlichkeit haploider und diploid-haploider Gemische von Eiern von Tetranychus urticae Koch gegenüber Tedion untersucht. Für einen normal empfindlichen Stamm wurde aus toxikologischen Daten und einer Verschiebung des Geschlechterverhältnisses erschlossen, daß haploide Eier gegenüber Tedion empfindlicher sind als diploide. Dieser Toleranzunterschied zwischen haploiden und diploiden Nachkommen konnte bei einem gegen Tedion resistenten Stamm nicht nachgewiesen werden.Es wurden Massenkreuzungen zwischen empfindlichen und resistenten Stämmen durch-geführt. Empfindliche Weibchen, mit resistenten Männchen gepaart, produzierten empfindliche haploide und resistente diploide Nachkommen. Resistente Weibchen, mit empfindlichen Männchen gepaart, ergaben eine resistente Nachkommenschaft. Beide Geschlechter können also die Resistenz gegen Tedion übertragen. Da bei den reziproken Kreuzungen ein Toleranzunterschied zwischen den diploiden Nachkommen auftritt, wird angenommen, daß in dem genetischen Mechanismus der Tedion-Resistenz auch eine mütterliche oder eine zytoplasmatische Komponente vorhanden ist.相似文献