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1.
Morgan Sheng Michael Wyszynski 《BioEssays : news and reviews in molecular, cellular and developmental biology》1997,19(10):847-853
Electrical signaling by neurons depends on the precisely ordered distribution of a wide variety of ion channels on the neuronal surface. The mechanisms underlying the targeting of particular classes of ion channels to specific subcellular sites are poorly understood. Recent studies have identified a new class of protein-protein interaction mediated by PDZ domains, protein binding modules that recognize specific sequences at the C terminus of membrane proteins. The PDZ domains of a family of synaptic cytoskeleton-associated proteins, typified by PSD-95, bind to the intracellular C-terminal tails of NMDA receptors and Shaker-type K+ channels. This interaction appears to be important in the clustering and localization of these ion channels at synaptic sites. Recognition of specific C-terminal peptide sequences by different PDZ domain-containing proteins may be a general mechanism for differential targeting of proteins to a variety of subcellular locations. 相似文献
2.
The cysteine conserved among DNA cytosine methylases is required for methyl transfer, but not for specific DNA binding. 总被引:9,自引:5,他引:4
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M W Wyszynski S Gabbara E A Kubareva E A Romanova T S Oretskaya E S Gromova Z A Shabarova A S Bhagwat 《Nucleic acids research》1993,21(2):295-301
All DNA (cytosine-5)-methyltransferases contain a single conserved cysteine. It has been proposed that this cysteine initiates catalysis by attacking the C6 of cytosine and thereby activating the normally inert C5 position. We show here that substitutions of this cysteine in the E. coli methylase M. EcoRII with either serine or tryptophan results in a complete loss of ability to transfer methyl groups to DNA. Interestingly, mutants with either serine or glycine substitution bind tightly to substrate DNA. These mutants resemble the wild-type enzyme in that their binding to substrate is not eliminated by the presence of non-specific DNA in the reaction, it is sensitive to methylation status of the substrate and is stimulated by an analog of the methyl donor. Hence the conserved cysteine is not essential for the specific stable binding of the enzyme to its substrate. However, substitution of the cysteine with the bulkier tryptophan does reduce DNA binding. We also report here a novel procedure for the synthesis of DNA containing 5-fluorocytosine. Further, we show that a DNA substrate for M. EcoRII in which the target cytosine is replaced by 5-fluorocytosine is a mechanism-based inhibitor of the enzyme and that it forms an irreversible complex with the enzyme. As expected, this modified substrate does not form irreversible complexes with the mutants. 相似文献
3.
Bañares VG Peterson G Aguilar D Gulayin R Sisu E Wyszynski DF Pivetta OH Tavella MJ 《Human biology; an international record of research》2005,77(2):247-256
Several studies have shown evidence of an association between the *4 allele of apolipoprotein E (APOE) and coronary heart disease (CHD) in different populations. We determined the APOE genotype and total cholesterol (TC), triglycerides (TG), and high-density lipoprotein cholesterol (HDLC) values in 189 patients with angiographically evaluated atherosclerosis. The APOE*4 allele was found to be statistically significantly more frequent (odds ratio, 1.93; 95% confidence interval, 1.12-3.32) among male patients than in a randomly chosen population-based sample. No significant difference was found when female patients were compared to the general population. The APOE*4 allele was found primarily among young (30-45-year-old) male patients (p < 0.04). Despite the ascending linear tendency of the mean TC values for genotypes APOE*2/*3, APOE*3/*3, and APOE*3/ *4 reported in our case population, no differences were observed among our patients. We conclude that the APOE*4 allele is associated with an increased risk for atherosclerotic vascular disease, that this association has an age-dependent effect, and that it acts as a genetic factor that increases susceptibility to developing the disease in young to middle-aged male adults in our population. 相似文献
4.
Numerous transposed sequences of mitochondrial cytochrome oxidase I-II in aphids of the genus Sitobion (Hemiptera: Aphididae) 总被引:4,自引:1,他引:3
Polymerase chain reaction (PCR) products corresponding to 803 bp of the
cytochrome oxidase subunits I and II region of mitochondrial DNA (mtDNA
COI-II) were deduced to consist of multiple haplotypes in three Sitobion
species. We investigated the molecular basis of these observations. PCR
products were cloned, and six clones from one individual per species were
sequenced. In each individual, one sequence was found commonly, but also
two or three divergent sequences were seen. The divergent sequences were
shown to be nonmitochondrial by sequencing from purified mtDNA and Southern
blotting experiments. All seven nonmitochondrial clones sequenced to
completion were unique. Nonmitochondrial sequences have a high proportion
of unique sites, and very few characters are shared between
nonmitochondrial clones to the exclusion of mtDNA. From these data, we
infer that fragments of mtDNA have been transposed separately (probably
into aphid chromosomes), at a frequency only known to be equalled in
humans. The transposition phenomenon appears to occur infrequently or not
at all in closely related genera and other aphids investigated. Patterns of
nucleotide substitution in mtDNA inferred over a parsimony tree are very
different from those in transposed sequences. Compared with mtDNA,
nonmitochondrial sequences have less codon position bias, more even
exchanges between A, G, C and T, and a higher proportion of nonsynonymous
replacements. Although these data are consistent with the transposed
sequences being under less constraint than mtDNA, changes in the
nonmitochondrial sequences are not random: there remains significant
position bias, and probable excesses of synonymous replacements and of
conservative inferred amino acid replacements. We conclude that a
proportion of the inferred change in the nonmitochondrial sequences
occurred before transposition. We believe that Sitobion aphids (and other
species exhibiting mtDNA transposition) may be important for studying the
molecular evolution of mtDNA and pseudogenes. However, our data highlight
the need to establish the true evolutionary relationships between sequences
in comparative investigations.
相似文献
5.
6.
Sam Gabbara Michael Wyszynski Ashok S. Bhagwat 《Molecular genetics and genomics : MGG》1994,243(2):244-248
Escherichia coli contains a base mismatch correction system called VSP repair that is known to correct T:G mismatches to C:G when they occur in certain sequence contexts. The preferred sequence context for this process is the site for methylation by the E. coli DNA cytosine methylase (Dcm). For this reason, VSP repair is thought to counteract potential mutagenic effects of deamination of 5-methylcytosine to thymine. We have developed a genetic reversion assay that quantitates the frequency of C to T mutations at Dcm sites and the removal of such mutations by DNA repair processes. Using this assay, we have studied the repair of U: G mismatches in DNA to C: G and have found that VSP repair is capable of correcting these mismatches. Although VSP repair substantially affects the reversion frequency, it may not be as efficient at correcting U: G mismatches as the uracil DNA glycosylase-mediated repair process. 相似文献
7.
Substitutions of a cysteine conserved among DNA cytosine methylases result in a variety of phenotypes. 总被引:14,自引:13,他引:1
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The proposed mechanism for DNA (cytosine-5)-methyltransferases envisions a key role for a cysteine residue. It is expected to form a covalent link with carbon 6 of the target cytosine, activating the normally inactive carbon 5 for methyl transfer. There is a single conserved cysteine among all DNA (cytosine-5)-methyltransferases making it the candidate nucleophile. We have changed this cysteine to other amino acids for the EcoRII methylase; which methylates the second cytosine in the sequence 5'-CCWGG-3'. Mutants were tested for their methyl transferring ability and for their ability to form covalent complexes with DNA. The latter property was tested indirectly with the use of a genetic assay involving sensitivity of cells to 5-azacytidine. Replacement of the conserved cysteine with glycine, valine, tryptophan or serine led to an apparent loss of methyl transferring ability. Interestingly, cells carrying the mutant with serine did show sensitivity to 5-azacytidine, suggesting the ability to link to DNA. Unexpectedly, substitution of the cysteine with glycine results in the inhibition of cell growth and the mutant allele can be maintained in the cells only when it is poorly expressed. These results suggest that the conserved cysteine in the EcoRII methylase is essential for methylase action and it may play more than one role in it. 相似文献
8.
Wyszynski DF 《Teratology》2001,64(4):221-225
This article enumerates the congenital anomalies mentioned in the Bible and the Talmud, the two holiest and oldest texts in Judaism. Most of these conditions were described to regulate attributes that would disqualify a Priest from performing religious rituals in the Temple in Jerusalem. However, the cultural atmosphere in Biblical and Talmudical times was one in which physical deformity did not necessarily evoke a negative aesthetic reaction, an assumption of ill health, or the expectation of economic dependence. 相似文献
9.
D F Wyszynski C T Baldwin M A Cleves Y Amirault V G Nolan J J Farrell A Bisbee A Kutlar L A Farrer M H Steinberg 《Cellular and molecular biology, including cyto-enzymology》2004,50(1):23-33
In patients with sickle cell anemia, fetal hemoglobin (HbF) concentrations vary by 2 orders of magnitude. This variance may be a result of heterogeneity in gene regulatory elements; accordingly, we searched for single nucleotide polymorphisms (SNPs) that might identify this variation. More than 180 SNPs were studied in 38 genes in 280 sickle cell anemia patients. The strongest association with HbF was found with SNPs near a QTL previously localized on chromosome 6q22.3-q23.2. Initially, two SNPs were identified in intergenic portions of this QTL and were associated with about a 20% difference in percent HbF. Subsequently, we genotyped 44 additional SNPs in the genomic region between 136.1 Mb and 137.5 Mb on chromosome 6q. Twelve SNPs, associated with a 20%-30% difference in HbF concentrations, were located in the introns of four genes, PDE7B, MAP7, MAP3K5 and PEX7. In K562 cells, the p38-MAPK pathway has been associated with the activation of gamma-globin gene expression by histone deacetylase inhibitors. Haplotypes C-T-T-T in MAP7 and T-C-C in PEX7 were significantly associated with increases in concentration of HbF, both showing strong dominance. Genetic elements abutting the 6q22.3-q23.2 QTL, may harbor trans-acting elements that help modulate baseline HbF level in sickle cell anemia. 相似文献
10.
We developed a new and simple method to collect sections of a whole brown rice kernel for investigation of histological properties. A single kernel of rice was dehydrated through a graded ethanol series, transferred to xylene, and embedded in paraffin. During sectioning of the blocks using a rotary microtome, we used a special adhesive tape to collect and place the sections on slides so they remained flat. The use of the adhesive tape technique combined with autofluorescence characteristics allowed us to visualize cell walls throughout an entire longitudinal or transverse section of a whole rice kernel. We obtained scanning electron microscopy images of the sections to determine section quality. 相似文献