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M Tone

Publications and source records attributed to M Tone.

26 records · Page 2Linked to original sources

Characterization of the CAMPATH-1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone.

The CAMPATH-1 (CDw52) antigen has been purified from human spleen. The antigenic epitope is heat stable but sensitive to mild alkali treatment. Experiments with phosphatidylinositol-specific phospholipase C indicate that it is anchored by a glycosylphosphatidylinositol (GPI) anchor. An N-terminal sequence of 11 amino acids was determined, followed by an abrupt stop. Using short overlapping mixed oligonucleotide primers, cDNA synthesized from the mRNA of a human B cell line was amplified by the polymerase chain reaction. The product was used to isolate cDNA clones and the full amino acid sequence of the CAMPATH-1 antigen was deduced. It consists of 37 amino acid residues plus a 24-residue signal peptide. It has all the features expected for a GPI-anchored membrane protein except that the predicted mature protein is remarkably short, comprising no more than 18 residues and possibly as few as 12 (depending on the GPI linkage site). Potential attachment sites for carbohydrate are present and it is shown that the antigen contains N-linked oligosaccharide(s). This structure accounts for the known properties of the antigen, though the exact reasons why it is such a good target for cell lysis in vitro and in vivo are not yet clear.

Amino Acid Sequence↗

[Clinical studies of 15 cases of progressive systemic sclerosis (PSS) associated with positive proteinuria and membranous glomerulonephritis].

In this study, we demonstrated that the patients with PSS who develop proteinuria during the course are much more prone to develop lung fibrosis, telangiectasia, serositis and esophageal hypomotility. They also exhibited positive LE cell phenomenon, lupus band test, thrombocytopenia and leukopenia. The mortality rate is significantly higher in proteinuria positive patients and abnormal clinical data described above seem to be the risk factors for the development of renal involvement in PSS.

Adult↗

Nucleotide sequences and unusual electrophoretic behavior of the W chromosome-specific repeating DNA units of the domestic fowl, Gallus gallus domesticus.

Nucleotide sequences of three independently cloned repeating units of the W chromosome-specific repetitive DNA sequences ("XhoI family") of the chicken were determined. All three units are 717 bp long with XhoI sites at both ends. There are only 21 sites out of 717 bases where a single base change occurs in one of the three clones. Each of these repeating units consists of 34 tandem repeats of about 21 bp. Sequences of some members of these internal repeats are not well conserved, but the majority of the repeats are characterized by the presence of (A)3-5 and (T)3-5 clusters separated by 6-7 relatively G + C-rich base pairs. One striking feature of the cloned 717 bp repeating units is that they migrate unusually slowly on electrophoresis in polyacrylamide gels. The same feature is also shown by a genomic population of the 0.7 kb repeating units recovered from XhoI digests of the genomic DNA of the female chicken. This anomalous behavior is attributed to the occurrence of DNA curvatures because of the above sequence characteristics and partial recovery of the electrophoretic mobility in the presence of distamycin A. Another feature of the 717 bp repeating unit is the presence of 438 and 159 nucleotide-long open reading frames (ORFs) at each end of the unit. A possible function of the XhoI family sequences in the heterochromatization of the W chromosome and the significance of the ORFs are discussed.

Animals↗

Structure of human alpha 2-plasmin inhibitor deduced from the cDNA sequence.

We have isolated three cDNA clones for human alpha 2-plasmin inhibitor (alpha 2-PI). Two clones are from human hepatoma cell line, Hep G2, and cover the entire protein coding region plus the 3'-flanking region up to the poly(A) sequence, and the other clone is from human liver and contains the carboxyl-terminal half. The total length of the cDNAs is 2.29 kb, corresponding to more than 95% of the full-length mRNA. alpha 2-PI seems to consist of 452 amino acid residues plus 39 amino acid residues for the signal peptide. The amino acid sequence shows 23 to 28% homology to those of five other protease inhibitors, plasminogen activator inhibitor (PAI), protein C inhibitor (PCI), alpha 1-antitrypsin (alpha 1-AT), antithrombin III (AT III), and alpha 1-antichymotrypsin (alpha 1-AC). alpha 2-PI seems to be the most distantly related among these inhibitors. Comparison of the phylogenetic trees of proteases and their inhibitors indicates that four proteases, namely elastase (or trypsin), chymotrypsin, plasminogen activator, and thrombin, may have evolved concurrently with the corresponding inhibitors. However, alpha 2-PI and PCI seem to have evolved asynchronously from their substrates. The data suggest that alpha 2-PI may originally have inhibited some protease other than plasmin, and protein C may have had an inhibitor different from the present one early in its evolutionary history.

Amino Acid Sequence↗

Genus specificity and extensive methylation of the W chromosome-specific repetitive DNA sequences from the domestic fowl, Gallus gallus domesticus.

Two female-specific repeating DNA units of 0.6 kilobase pairs (kb) and 1.1 kb, produced by digesting the genomic DNA of the White Leghorn chicken with Xho I, were cloned by inserting them into the Xho I site of an Escherichia coli plasmid vector pACYC177. Two such recombinant plasmids, pAGD0601 and pAGD1101, containing a single 0.6-kb and 1.1-kb sequence, respectively, were used as molecular probes. In situ hybridization of the 3H-probes to the metaphase chromosomes from the female White Leghorn embryos revealed their localization in the W chromosome. Semiquantitative Southern blot hybridization with 32P-probes in excess indicated that the 0.6-kb unit and 1.1-kb unit were repeated approximately 14,000 and 6,000 times, respectively, in the W chromosome. The two units comprised about 46% of the W chromosomal DNA. These two repeating units were found in the female genomes of every line of Gallus g. domesticus tested and in the female genomes of three jungle fowl species (G. gallus, G. sonneratii, and G. varius) but not in three species belonging to other genera in the suborder Galli. Hha I sites in the 0.6-kb and 1.1-kb repeating units were shown to be extensively methylated and a significant fraction of the Hpa II sites in the 0.6-kb repeating units were also shown to be methylated in the female genome of the White Leghorn. Methylation patterns of Hpa II sites in or around the 0.6-kb repeating units examined by the Msp I digestion were similar in the various lines of domestic fowls and the two species of jungle fowls, but G varius (black or green jungle fowl) produced a different pattern of digestion with Msp I.

Animals↗

Demonstration of W chromosome-specific repetitive DNA sequences in the domestic fowl, Gallus g. domesticus.

Evidence is presented to demonstrate the presence of W chromosome-specific repetitive DNA sequences in the female White Leghorn chicken, Gallus g. domesticus, based on two different experimental approaches. First, 3H-labelled, female chicken DNA was hybridized with excess, unlabelled, mercurated, male DNA, and unhybridized single-stranded 3H-DNA (3H-SHU-DNA) was recovered by SH-Sepharose and hydroxyapatite column chromatography. Approximately 24% of the hybridizable 3H-SHU-DNA was female-specific and localized on the W chromosome. The second approach was to examine female-specific DNA fragments among the digests of chicken DNA with various restriction endonucleases. Among them, we found that digestion with XhoI produced two prominent female-specific brands of 0.60 kb (= kilobase pairs) and 1.1 kb. The 0.60 kb fragment was isolated and 3H-labelled by nick-translation. Female-specificity of the 3H-XhoI--0.60 kb DNA was judged to be at least 95% under the conditions of hybridization with membrane filter-bound DNA. Presence of amplified XhoI--0.60 kb DNA on the W chromosome seems to be limited to different lines of G.g. domesticus and no such repeat was detected in three species belonging to other genera in the order Galliformes and in three species belonging to other avian orders.

Animals↗

Variation of repetitive DNA and its phylogenetic relation in Hynobiidae (Caudata).

Middle repetitive (MR) DNA comprising about 35% of the genomic DNA of Hynobius retardatus was isolated and used as mixed probes. A 1.3-kb Bam HI fragment, the major repeating unit of the extensively amplified sequences in H. retardatus, was cloned, sequenced, and used as a probe. Dot blot hybridization under the conditions of probe excess revealed that the MR DNA sequences were shared among the three salamander genera belonging to the family Hynobiidae. However, a large difference in the amount of common sequences was found between Hynobius and Salamandrella, and large differences in both amounts and sequence similarity were found between Hynobius and Onychodactylus. Even among species belonging to the genus Hynobius, relative contents of sequences common to the H. retardatus MR DNA were 20% to 50%. Partially related sequences to the 1.3-kb sequence seemed to be present in all the species of Hynobius and Salamandrella, but the extensive amplification was found only in H. retardatus. Based on the relative contents of the common MR DNA and previously reported chromosomal banding characteristics, species in the genus Hynobius are classified into at least three groups. An evolutionary aspect of the emergence of the amplified sequence in H. retardatus is discussed.

Animals↗