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The Saccharomyces cerevisiae CKS1 gene, a homolog of the Schizosaccharomyces pombe suc1+ gene, encodes a subunit of the Cdc28 protein kinase complex. 总被引:22,自引:7,他引:15
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J A Hadwiger C Wittenberg M D Mendenhall S I Reed 《Molecular and cellular biology》1989,9(5):2034-2041
The Saccharomyces cerevisiae gene CDC28 encodes a protein kinase required for cell cycle initiation. In an attempt to identify genes encoding proteins that interact with the Cdc28 protein kinase, high-copy plasmid suppressors of a temperature-sensitive cdc28 mutation were isolated. One such suppressor, CKS1, was found to encode an 18-kilodalton protein that shared a high degree of homology with the suc1+ protein (p13) of Schizosaccharomyces pombe (67% amino acid sequence identity). Disruption of the chromosomal CKS1 gene conferred a G1 arrest phenotype similar to that of cdc28 mutants. The presence of the 18-kilodalton Cks1 protein in yeast lysates was demonstrated by using Cks-1 specific antiserum. Furthermore, the Cks1 protein was shown to be physically associated with active forms of the Cdc28 protein kinase. These data suggest that Cks1 is an essential component of the Cdc28 protein kinase complex. 相似文献
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The disease resistance response in pea is associated with increased levels of specific mRNAs 总被引:1,自引:1,他引:0
A cDNA library was constructed using poly(A)+RNA fromPisum sativum which had been treated for 8 h with the fungusFusarium solani f. sp.phaseoli. Two thousand four hundred recombinant colonies were screened by differential colony hybridization using32P-labelled cDNAs prepared from RNA extracted from either noninoculated or inoculated pea tissue. cDNA clones were then selected,
which showed greater hybridization with cDNA prepared from pea RNA 8 h post-inoculation than with a cDNA probe from 0 h. Seven
distinct hybridization classes were chosen for further study. Northern blot analyses of total cellular RNAs inoculated for
16 h with eitherF. solani phaseoli or water demonstrated that each cDNA clone selected represents an mRNA species which increases substantially in abundance
during infection.
Results of3H-uridine pulse-labelling experiments suggested that enhanced synthesis is at least partially responsible for the accumulation
of the fungus-inducible mRNAs which hybridized with the clones. 相似文献
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Isolation and characterization of a large,neurite-associated glycoconjugate from neuroblastoma cells
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A high molecular weight glycoconjugate has been isolated from neurite-producing neuronal tumor cells in culture and has been designated as I(0) based on its elution characteristics in gel filtration chromatography. This molecule cannot be found in a variety of nonneuronal cells. I(0) is found in the substratum-attached material or cell fraction of neurite-producing neuroblastoma cells, depending upon culture conditions. It is found in the substratum-bound fraction of B104 rat neuroblastoma cells during serum starvation and in the EGTA-detached cell fraction of B104 cells grown in chemically defined N2 medium. It occurs only in the cell fraction of the human neuroblastoma line Platt. Examination of behavioral variants of the B104 rat line further strengthens the association of I(0) with neurite production; the constitutive neurite-producing E(R)B9 variant contains I(0) while the non-neurite-producing E(R)A11 variant does not. I(0) is large, eluting in the void volume of sepharose-CL2B columns. Radioiodination of intact cells with lactoperoxidase shows I(0) to be a cell surface component. Metabolic radiolabeling studies show that it contains a high proportion of polysaccharide to protein, does not contain mannose, and is unsulfated. Alkaline borohydride reduction release two size classes of large polysaccharide chain. The alkaline reduction results, along with the mannose incorporation studies, show the presence of O-glycosidic linkages and few, if any, N-linkages. Resistance to nitrous acid deamination, insensitivity to glycosaminoglycan lyases, and the absence of sulfation, indicate that I(0) does not contain the glycosaminoglycans hyaluronic acid, chondroitin-, dermatan-, or heparin- sulfates. Affinity column chromatography reveals high binding affinity of I(0) to polyornithine and no binding to gelatin (collagen) or the glycosaminoglycans hyaluronate and heparin. These studies describe a unique high molecular weight glycoconjugate on the surface of neurite-producing neuroblastoma cell lines from two species. 相似文献
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Deep-level diagnostic value of the rDNA-ITS region 总被引:14,自引:0,他引:14
The similarity of certain reported angiosperm rDNA internal transcribed
spacer (ITS) region sequences to those of green algae prompted our analysis
of the deep-level phylogenetic signal in the highly conserved but short
5.8S and hypervariable ITS2 sequences. We found that 5.8S sequences yield
phylogenetic trees similar to but less well supported than those generated
by a ca. 10-fold longer alignment from rDNA-18S sequences, as well as
independent evidence. We attribute this result to our finding that,
compared to 18S, the 5.8S has a higher proportion of sites subject to vary
and greater among-site substitution rate homogeneity. We also determined
that our phylogenetic results are not likely affected by intramolecular
compensatory mutation to maintain RNA secondary structure nor by evident
systematic biases in base composition. Despite historical homology, there
appears to be no ITS2 primary sequence similarity shared sufficient
similarity to cluster correctly on the basis of alignability. Our results
indicate that groups, however, share sufficient similarity to cluster
correctly on the basis of alignability. Our results indicate that ITS
region sequences can diagnose organismal origins and phylogenetic
relationships at many phylogenetic levels and provide a useful paradigm for
molecular evolutionary study.
相似文献
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Production of Serine Proteases by the Oyster Pathogen Perkinsus marinus (Apicomplexa) In Vitro 总被引:1,自引:1,他引:0
JEROME F. LA PEYRE DORIS Y. SCHAFHAUSER ESAM H. RIZKALLA MOHAMED FAISAL 《The Journal of eukaryotic microbiology》1995,42(5):544-551
ABSTRACT. Analysis of the cell-free supernatants of Perkinsus marinus cultures by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and silver staining revealed the presence of as many as 17 bands ranging in molecular weight from 239 to 32 kDa. These bands were not present in un-inoculated medium. Moreover, P. marinus produces extracellular proteins that possess proteolytic activities; the cell-free supernatants of P. marinus cultures could digest a variety of proteins including gelatin, casein, fibronectin and laminin. Oyster plasma was also digested by cell-free culture supernatants. The proteolytic activity in cell-free culture supernatants was detected 24 h post-inoculation, while no proteolytic activity could be detected in cell lysates. The proteolytic activities were characterized using substrate-impregnated sodium dodecylsulfate-polyacrylamide gels and had approximate molecular weights ranging from 55 to 35 kDa. The proteolytic activity of cell-free culture supernatants was inhibited by the serine protease inhibitors phenylmethylsulphonyl fluoride, 3,4-dichloroisocoumarin and soybean trypsin inhibitor. In contrast, inhibitors (i.e. trans-epoxysuccinyll-leucylamido(4-guanidino)-butane, 1, 10-phenanthroline, captopril, ethylenediaminetetracetic acid, pepstatin A or diazoacetyl-DL-norleucine methyl ester) from the other three classes of proteases had no effect. It was concluded that the P. marinus proteases in cell-free culture supernatants are serine proteases. 相似文献
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