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101.
L. Hendriks R. De Baere Y. Van de Peer J. Neefs A. Goris R. De Wachter 《Journal of molecular evolution》1991,32(2):167-177
Summary The complete small ribosomal subunit RNA (srRNA) sequence was determined for the red algaPorphyra umbilicalis and the basidiomyceteLeucosporidium scottii, representing two taxa for which no srRNA sequences were hitherto known. These sequences were aligned with other published
complete srRNA sequences of 58 eukaryotes. Evolutionary trees were reconstructed by a matrix optimization method from a dissimilarity
matrix based on sections of the alignment that correspond to structurally conservative areas of the molecule that can be aligned
unambiguously. The overall topology of the eukaryotic tree thus constructed is as follows: first there is a succession of
early diverging branches, leading to a diplomonad, a microsporidian, a euglenoid plus kinetoplastids, an amoeba, and slime
molds. Later, a nearly simultaneous radiation seems to occur into a number of taxa comprising the metazoa, the red alga, the
sporozoa, the higher fungi, the ciliates, the green plants, plus some other less numerous groups. Because the red alga diverges
late in the evolutionary tree, it does not seem to represent a very primitive organism as proposed on the basis of morphological
and 5S rRNA sequence data.
Asco- and basidiomycetes do not share a common ancestor in our tree as is generally accepted on the basis of conventional
criteria. In contrast, when all alignment positions, rather than the more conservative ones, are used to construct the evolutionary
tree, higher fungi do form a monophyletic cluster. The hypothesis that higher fungi and red algae might have shared a common
origin has been put forward. Although the red alga and fungi seem to diverge at nearly the same time, no such relationship
can be detected.
The newly determined sequences can be fitted into a secondary structure model for srRNA, which is now relatively well established
with the exception of uncertainties in a number of eukaryote-specific expansion areas. A specific structural model featuring
a pseudoknot is proposed for one of these areas. 相似文献
102.
Differential methylation at the 5′ and the 3′ CCGG sites flanking the X chromosomal hypervariable DXS255 locus 总被引:3,自引:0,他引:3
R. W. Hendriks M. E. M. Kraakman R. G. J. Mensink R. K. B. Schuurman 《Human genetics》1991,88(1):105-111
Summary The degree of methylation at the 5 and 3 CCGG sequences flanking the variable number of tandem repeat (VNTR) region of the DXS255 locus at Xp11.22 was analysed separately in several haematopoietic cell lineages. The 5 CCGG site on active chromosomes was found to be completely methylated in B and T lymphocytes and granulocytes. Methylation of the 5 site on inactive X chromosomes differed between females (0%–60%), but was consistent in different cell lineages obtained from individual females. In contrast, methylation at the 3 CCGG site on active chromosomes was found to vary in B lymphocytes (40%–100%), whereas complete methylation was found in T lymphocytes and granulocytes. The extent of methylation on inactive X chromosomes was found to differ significantly between B lymphocytes (17%), T lymphocytes (54%) and granulocytes (82%). Thus, methylation at the 5 CCGG site seems to be primarily related to the status of X chromosome inactivation, whereas methylation at the 3 CCGG site is mainly subject to cell-lineage-specific influences. 相似文献
103.
Jos L. M. Lebouille Rudi W. Hendriks Nell M. Soeter J. Peter H. Burbach 《Journal of neurochemistry》1989,52(6):1714-1721
Incubation of beta-endorphin with cytosolic and particulate fractions of rat brain resulted in the formation of several peptides, including gamma-endorphin [beta-endorphin-(1-17)] and beta-endorphin-(18-31), indicating the presence of enzyme activity cleaving the Leu17-Phe18 bond of beta-endorphin. An assay for this Leu-Phe cleaving activity, based on the cleavage of the 14C-labeled substrate acetyl-Val-Thr-Leu-Phe-[epsilon-([14C]CH3)2]Lys-NHCH3, was used to examine the properties of this enzyme activity. beta-Endorphin-(1-31) competitively inhibited the Leu-Phe-cleaving enzyme activity on the pentapeptide substrate. Over 90% of activity was recovered in the cytosolic fraction. Leu-Phe-cleaving activity behaved like a thiol endopeptidase because it was inhibited by low concentrations of N-ethylmaleimide, p-chloromercuribenzoate, p-chloromercuribenzoyl sulfate, and low concentrations of Hg2+. Low concentrations of sulfhydryl compounds stimulated Leu-Phe-cleaving activity. The activity was optimal between pH 8.5 and 9.0. The Km of Leu-Phe-cleaving activity in the cytosolic fraction was 35 microM and in the particulate fraction 88 microM with Vmax values of 193 and 15 nmol mg protein-1 h-1, respectively. The apparent molecular mass of the Leu-Phe-cleaving enzyme was estimated by gel filtration to be approximately 200 kilodaltons. These properties of Leu-Phe-cleaving activity indicate that the Leu-Phe-cleaving enzyme is distinct from any known brain endopeptidase. 相似文献
104.
105.
Primary and secondary structure of the 18 S ribosomal RNA of the insect species Tenebrio molitor 总被引:5,自引:0,他引:5
The sequence of the 18 S rRNA of Tenebrio molitor is reported. A detailed secondary structure model for eukaryotic small subunit rRNAs is proposed. The model comprises 48 universal helices that eukaryotic and prokaryotic small subunit rRNAs have in common, plus a number of helices in areas of variable secondary structure. For the central area of the model, an alternative structure is possible, applicable only to eukaryotic small subunit rRNAs. Possibly, small subunit rRNA switched to this alternative conformation after the eukaryotic branch had been established in evolution. Another possibility is that the two conformers represent a dynamic structural switch functioning during the translational activity of the eukaryotic ribosome. 相似文献
106.
107.
In the active centre of pancreatic phospholipase A2 His48 is at hydrogen-bonding distance to Asp99. This Asp-His couple is assumed to act together with a water molecule as a catalytic triad. Asp99 is also linked via an extended hydrogen bonding system to the side chains of Tyr52 and Tyr73. To probe the function of the fully conserved Asp99, Tyr52 and Tyr73 residues in phospholipase A2, the Asp99 residue was replaced by Asn, and each of the two tyrosines was separately replaced by either a Phe or a Gln. The catalytic and binding properties of the Phe52 and Phe73 mutants did not change significantly relative to the wild-type enzyme. This rules out the possibility that either one of the two Tyr residues in the wild-type enzyme can function as an acyl acceptor or proton donor in catalysis. The Gln73 mutant could not be obtained in any significant amounts probably due to incorrect folding. The Gln52 mutant was isolated in low yield. This mutant showed a large decrease in catalytic activity while its substrate binding was nearly unchanged. The results suggest a structural role rather than a catalytic function of Tyr52 and Tyr73. Substitution of asparagine for aspartate hardly affects the binding constants for both monomeric and micellar substrate analogues. Kinetic characterization revealed that the Asn99 mutant has retained no less than 65% of its enzymatic activity on the monomeric substrate rac 1,2-dihexanoyldithio-propyl-3-phosphocholine, probably due to the fact that during hydrolysis of monomeric substrate by phospholipase A2 proton transfer is not the rate-limiting step.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
108.
109.