Lichen-forming fungi, in symbiotic associations with algae, frequently have
nuclear small subunit ribosomal DNA (SSU rDNA) longer than the 1,800
nucleotides typical for eukaryotes. The lichen-forming ascomycetous fungus
Lecanora dispersa contains insertions at eight distinct positions of its
SSU rDNA; the lichen-forming fungi Calicium tricolor and Porpidia
crustulata each contain one insertion. Insertions are not limited to fungi
that form lichens; the lichen ally Mycocalicium albonigrum also contains
two insertions. Of the 11 insertion positions now reported for
lichen-forming fungi and this ally, 6 positions are known only from
lichen-forming fungi. Including the 4 newly reported in this study,
insertions are now known from at least 17 positions among all reported SSU
rDNA sequences. Insertions, most of which are Group I introns, are reported
in fungal and protistan lineages and occur at corresponding positions in
genomes as phylogenetically distant as the nuclei of fungi, green algae,
and red algae. Many of these positions are exposed in the mature rRNA
tertiary structure and may be subject to independent insertion of introns.
Insertion of introns, accompanied by their sporadic loss, accounts for the
scattered distribution of insertions observed within the SSU rDNA of these
diverse organisms.
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The increased polylactosamine glycosylation of LAMP-2 in MDCK cells
cultured for 1 day relative to cells cultured for 3 days has been
correlated with its slower rate of Golgi transit (Nabi and Rodriguez-
Boulan, 1993, Mol. Biol. Cell., 4, 627-635). To determine if the
differential polylactosamine glycosylation of LAMP-2 is a consequence of
glycosyltransferase expression levels, the activities of beta1- 6GlcNAc-TV,
beta1-3GlcNAc-T(i), beta1-2GlcNAc-TI, beta1, 4Gal-T, alpha2- 6sialyl-T, and
alpha2-3sialyl-T were assayed and no significant differences in the
activities of these enzymes in 1 and 3 day cell extracts were detected.
During MDCK epithelial polarization, the Golgi apparatus undergoes
morphological changes and apiconuclear Golgi networks were more evident in
3 day cells. Treatment with nocodazole disrupted Golgi networks and
generated numerous Golgi clusters in both 1 day and 3 day cells. In the
presence of nocodazole the differential migration of LAMP-2 in 1 and 3 day
MDCK cells was maintained and could be eliminated by treatment with
endo-beta-galactosidase, indicating that gross Golgi morphology did not
influence the extent of LAMP-2 polylactosamine glycosylation. Nocodazole
treatment did, however, result in the faster migration of LAMP-2 which was
not due to modification of core N-glycans as the precursor form of the
glycoprotein migrated with an identical molecular size. Following
incubation at 20 degrees C, which prevents the exit of proteins from the
trans-Golgi network, the molecular size of LAMP-2 increased to a similar
extent in both 1 and 3 day MDCK cells. Extending the time of incubation at
20 degrees C did not influence the size of LAMP-2, demonstrating that its
glycosylation is modified not by its retention within the Golgi but rather
by its equivalent slower Golgi passage at the lower temperature in both 1
and 3 day cells. An identical effect was observed in nocodazole treated
cells, demonstrating that Golgi residence time determines the extent of
LAMP-2 polylactosamine glycosylation, even in isolated Golgi clusters.
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An enzyme extract from the phycocyanin-containing unicellular rhodophyte, Cyanidium caldarium, reductively transforms biliverdin IX alpha to phycocyanobilin, the chromophore of phycocyanin, in the presence of NADPH. Unpurified cell extract forms both 3(E)-phycocyanobilin, which is identical to the major pigment that is released from phycocyanin by methanolysis, and 3(Z)-phycocyanobilin, which is obtained as a minor methanolysis product. After removal of low molecular weight material from the cell extract, only 3(Z)-phycocyanobilin is formed. 3(E)-Phycocyanobilin formation from biliverdin IX alpha, and the ability to isomerize 3(Z)-phycocyanobilin to 3(E)-phycocyanobilin, are reconstituted by the addition of glutathione to the incubation mixture. Partially purified protein fractions derived from the initial enzyme extract form 3(Z)-phycocyanobilin plus two additional, violet colored bilins, upon incubation with NADPH and biliverdin IX alpha. Further purified protein fractions produce only the violet colored bilins from biliverdin IX alpha. One of these bilins was identified as 3(Z)-phycoerythrobilin by comparative spectrophotometry, reverse-phase high pressure liquid chromatography, and 1H NMR spectroscopy. A C. caldarium protein fraction catalyzes the conversion of 3(Z)-phycoerythrobilin to 3(Z)-phycocyanobilin. This fraction also catalyzes the conversion of 3(E)-phycoerythrobilin to 3(E)-phycocyanobilin. The conversion of phycoerythrobilins to phycocyanobilins requires neither biliverdin nor NADPH. The synthesis of phycoerythrobilin and its conversion to phycocyanobilin by extracts of C. caldarium, a species that does not contain phycoerythrin, indicates that phycoerythrobilin is a biosynthetic precursor to phycocyanobilin. The enzymatic conversion of the ethylidine group from the Z to the E configuration suggests that the E-isomer is the precursor to the protein-bound chromophore. 相似文献
Factors favouring pollen callus proliferation, induction of embryogenesis and plant regeneration from cultured anthers of Digitalis obscura L. were determined. The presence of auxins was essential for cell proliferation and morphogenesis, and incubation in darkness singificantlyincreased these responses. Callus proliferation usually preceded embryo development, although sometimes direct embryogenesis was observed. On the other hand, bud differentiation was achieved only when callus was transferred to media containing cytokinin or several auxin/cytokinin combinations. Different ploidy levels] were observed in the regenerated plants, with approximately 50% being haploid. 相似文献
Plant dehydration is commonly caused by some adverse environmental conditions such as salinity, drought and freezing. As the plant hormone abscisic acid (ABA) is involved in responses to water stress, we studied its putative relationship with the degree of tolerance to these abiotic stresses. For this purpose we used cell lines that had been established from mature embryos of rice (Oryza sativa L, cvs. Bahia and Bomba), and selected by their high (L-T) or low (L-S) levels of tolerance to each type of stress. Tolerance of rice calli to either osmotic, saline, or freezing stress was generally improved by a previous treatment with ABA. This ABA effect was evident in those callus lines with low tolerance (L-S), as their ability to recover from stress increased up to three fold. Independent of the cultivar used, there were no significant differences in the endogenous ABA contents between untreated L-T and L-S lines. However, upon stress, the increase in endogenous ABA was higher in L-T than in L-S lines. These results, together with those obtained by using Fluridone, an inhibitor of ABA synthesis, show that differences in the level of cell tolerance to osmotic, saline and freezing stress are related to their different capacity of ABA synthesis under stress conditions. 相似文献
Cell-free extracts of the unicellular red alga Cyanidium caldarium catalyze the transformation of biliverdin to a product indistinguishable from phycocyanobilin, the free bilin derived from phycocyanin by methanolysis. Crude cell-free extract requires biliverdin as the only substrate, but after removal of low molecular weight components by gel filtration, the reaction shows an additional requirement for a reduced pyridine nucleotide. Boiled extract is enzymically inactive, activity is not sedimented by high-speed centrifugation, and mesobiliverdin cannot serve as a substrate.
Incubation of cell extracts with biliverdin yields two products with very similar spectrophotometric properties in acidic methanol, but which are separable by reverse-phase high pressure liquid chromatography. The same two products are formed by methanolysis of protein-bound phycocyanin chromophore, with the late-eluting one predominating. The two products derived from either phycocyanin methanolysis or cell extract incubation with biliverdin are partially interconvertible and they form the same ethylidine-free isomeric derivative, mesobiliverdin. Their absorption spectra correspond to those of the Z- and E-ethylidine isomers of phycocyanobilin. Based on previous work showing that the major methanolysis product has the E-ethylidine configuration, the other product of methanolysis and enzymic biliverdin transformation is therefore the Z-ethylidine isomer. The time course for formation of the two products during incubation suggests that the early-eluting product is the precursor of the late-eluting one. These results suggest that Z-ethylidine phycocyanobilin is the precursor of the E-ethylidine isomer, and that the latter may be a normal cellular precursor to protein-bound phycocyanin chromophore.
The Schwann cells are the myelinating glia of the peripheral nervous system that originated during development from the highly
motile neural crest. However, we do not know what the guidance signals for the Schwann cell precursors are. Therefore, we
set to test some of the known neurotrophins that are expressed early in developing embryos and have been shown to be critical
for the survival and patterning of developing glia and neurons. The goal of this study was to determine more specifically
if GDNF, NRG1 and NGF are chemoattractants and/or chemokinetic molecules for a Schwann cell precursor line, the Spl201. We
performed live chemoattraction assays, with imaging and also presented these molecules as part of their growing substrate.
Our results show for the first time that GDNF and NRG1 are potent chemoattractive and chemokinetic molecules for these cells
while NGF is a chemokinetic molecule stimulating their motility. 相似文献