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Biomedical subjects

R Yuan

Publications and source records attributed to R Yuan.

At least 73 records · Page 4Linked to original sources
PubMed1991

Isolation of temperature-sensitive McrA and McrB mutations and complementation analysis of the McrBC region of Escherichia coli K-12.

We isolated temperature-sensitive mcrA and mcrBC mutants of Escherichia coli. At 42 degrees C, they were unable to restrict the T-even bacteriophages T6gt and T4gt or plasmids encoding cloned DNA methylase genes whose specificities confer sensitivity to the McrA and McrBC nucleases. Complementation analysis of the McrBC region (mcrB251) with the complete cloned McrBC system or a derivative with mcrB alone indicated that the mutation shows an absolute defect for the restriction of DNA containing hydroxymethylcytosine and a thermosensitive defect for the restriction of DNA containing methylcytosine. The properties of the McrA temperature-sensitive mutants suggest that some of these mutations can also influence the restriction of DNA containing hydroxymethylcytosine or methylcytosine residues.

Chromosomes, Bacterial↗
PubMed1989

Activation of HIV LTR-directed expression: analysis with pseudorabies virus immediate early gene.

The long terminal repeat (LTR) region of the human immunodeficiency virus (HIV-1), which regulates viral gene expression, is modulated by viral trans-acting proteins of HIV and DNA viruses and by biologically active chemical agents that induce cellular proliferation and/or differentiation. The pseudorabies virus immediate early gene (PIE) shares similar transcriptional trans-activating properties with the gene products of several other DNA viruses. The transient expression chloramphenicol acetyl transferase (CAT) assays in HeLa cells transfected with HIV long terminal repeat (LTR)-CAT and PIE plasmids demonstrated trans-activation of the HIV LTR by PIE. Analyses of 5' deletion mutants and site-directed Sp1 and transactivation responsive (TAR) region mutants of the LTR indicated PIE-responsive sequences located between -65 and -17. Synergistic cooperativity between PIE and the HIV-1 tat protein was demonstrated. PIE exhibited a marked stimulatory effect upon HIV replication in HeLa cells transfected with a biologically active HIV proviral DNA. These data provide evidence that, like a number of other DNA containing viruses, PRV can trans-activate HIV gene expression.

Chromosome Mapping↗
PubMed1988

Neisseria gonorrhoeae M.Ngo AI DNA methyltransferase: physical and catalytic properties of the homogeneous enzyme.

A DNA methyltransferase, M.NgoAI, was purified to homogeneity from Neisseria gonorrhoeae strain WR220 by successive column chromatography. Its Mr is 25,000, as determined by both gel filtration and denaturing polyacrylamide gel electrophoresis. Maximal enzymatic activity was obtained in 50 mM Tris.HCl (pH 7.4), 10 mM EDTA, with incubation at 37 degrees C. An apparent Km value for S-adenosylmethionine and 5' -GGCC sites was determined to be 1.25 microM and 89.6 nM, respectively.

Bacterial Proteins↗
PubMed1988

Purification and characterization of DNA methyltransferases from Neisseria gonorrhoeae.

Three DNA methyltransferases, M.NgoAI, and M.NgoBI and M.NgoBII, free of any nuclease activities were isolated from Neisseria gonorrhoeae strains WR220 and MUG116 respectively. M.NgoAI recognizes the sequence 5' GGCC 3' and methylates the first 5' cytosine on both strands. M.NgoBI and M.NgoBII recognize 5' TCACC 3' and 5' GTAN5CTC 3' respectively. M.NgoBII methylates cytosine on only one strand to produce 5' GTAN5mCTC 3'.

Cytosine↗
PubMed1985

The characteristics of DNA methylation in an in vitro DNA synthesizing system from mouse fibroblasts.

An in vitro DNA synthesizing system from mouse fibroblasts has been used to study DNA methylation. DNA methylation occurs in two phases, one at the replication fork and the other farther behind it. Although 4% of the dCMP residues in mouse cell DNA are mdCMP, only 1.7% of the total [alpha 32P]dCMP in newly replicated DNA is methylated in vitro. No methylation of Okazaki fragments was detected. Nearest neighbor analysis of the newly replicated DNA revealed that, although 40% of the CpG dinucleotides were methylated, significant amounts of cytosine methylation were also found in CpC, CpT, and CpA dinucleotides.

Animals↗
PubMed1985

Inhibition of the type I restriction-modification enzymes EcoB and EcoK by the gene 0.3 protein of bacteriophage T7.

The gene 0.3 protein of bacteriophage T7 is a potent inhibitor of the restriction-modification enzymes EcoB and EcoK, both in vivo and in vitro. We have analyzed the ability of purified 0.3 protein to inhibit different steps in the reactions of EcoB and EcoK with DNA. Most of our experiments were done with EcoK, but selected tests with EcoB indicate that the two enzymes are affected by 0.3 protein in the same way. Purified 0.3 protein binds tightly to free enzyme, apparently to one of the small subunits, and prevents it from binding to DNA. If EcoK is allowed to form specific recognition complexes with unmodified DNA before 0.3 protein is added, relatively low levels of 0.3 protein prevent the nuclease activity that would otherwise appear upon addition of ATP, but considerably higher levels are needed to prevent formation of filter-binding complexes or ATPase activity. This, together with other results, suggests that the binding site for 0.3 protein is protected in recognition complexes and in the early stages of the ATP-stimulated reactions, but that it becomes accessible again before cleavage of the DNA, perhaps after the translocation step. If added after the nuclease reaction is substantially over, 0.3 protein has little effect on ATPase activity, and indeed, the subunit having the binding site for 0.3 protein apparently dissociates from the enzyme-DNA complex. The methylase activity of EcoK on hemi-methylated recognition sites is strongly inhibited by 0.3 protein added at any stage of the reaction.

Adenosine Triphosphate↗
PubMed1982

Rupture of a previously unruptured giant middle cerebral artery aneurysm after extracranial-intracranial bypass surgery.

The fatal rupture of a previously unruptured giant middle cerebral artery aneurysm occurred 13 days after an extracranial-intracranial bypass had been carried out, before definitive aneurysm surgery. Alterations in blood flow adjacent to the aneurysm after the bypass may have led to the fatal hemorrhage. After a preliminary extracranial-intracranial bypass procedure, there should be no undue delay in the direct attack on a giant aneurysm, regardless of its mode of presentation.

Cerebral Revascularization↗
PubMed1981

Characterization of the DNA methylase activity of the restriction enzyme from Escherichia coli K.

The restriction endonuclease from Escherichia coli K is a multifunctional protein which efficiently methylates heteroduplex DNA (one strand modified and one strand unmodified) in the presence of S-adenosylmethionine (AdoMet), ATP, and Mg2+. The methylase activity is catalytic, and seems to modify different heteroduplex host specificity sites for E. coli K with equal efficiency. In the methylase reaction, both AdoMet and ATP (or its imido analog) act as allosteric effectors, but AdoMet also serves as a methyl donor. Preincubation of the enzyme with AdoMet eliminates the lag period observed in DNA methylation. The rate of enzyme activation was determined using the AdoMet analog Sinefungin. The result are consistent with the hypothesis that the early steps of AdoMet binding and enzyme activation are common to both restriction and modification reactions.

Bacteriophage lambda↗