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L Snyder

Publications and source records attributed to L Snyder.

At least 73 records · Page 4Linked to original sources
PubMed1986

Molecular proof that bacteriophage T4 alc and unf genes are the same gene.

The DNA of bacteriophage T4 normally has a substituted base, hydroxymethylcytosine, instead of the usual cytosine. The bacteriophage shuts off host transcription after infection presumably by specifically blocking transcription of cytosine DNA. If T4 incorporates cytosine into its own DNA, this shutoff mechanism is directed back at itself and blocks its own transcription. Mutations which overcome this transcriptional block are in the T4 alc gene, and alc mutations allow the propagation of T4 with cytosine in their DNA (L. Snyder, L. Gold, and E. Kutter, Proc. Natl. Acad. Sci. USA 73:3098-3102, 1976). By genetic criteria, alc is the same as another gene, unf, whose product is required for the unfolding of the bacterial nucleoid after infection (K. Sirotkin, J. Wei, and L. Snyder, Nature [London] 265:28-32, 1977; D. P. Snustad, M. A. Tigges, K. A. Parson, C. J. H. Bursch, F. M. Caron, J. F. Koerner, and D. J. Tutas, J. Virol. 17:622-641, 1976; M. Tigges, C. J. H. Bursch, and D. P. Snustad, J. Virol. 24:775-785, 1977). The product of the alc gene has been identified as a 19-kilodalton protein (R. E. Herman, N. Haas, and D. P. Snustad, Genetics 108:305-317, 1984; E. Kutter, R. Drivdahl, and K. Rand, Genetics 108:291-304, 1984), and an open reading frame has been proposed to be the alc gene based on its size and map position (E. Kutter, R. Drivdahl, and K. Rand, Genetics 108:291-304, 1984). We used marker rescue techniques and DNA sequencing to confirm that this open reading frame is the alc gene. We also present a molecular proof that alc and unf are the same gene. While these results do not rigorously exclude the possibility that Unf and Alc are different activities of the same protein, they strongly support the conclusion that the unfolding of the bacterial nucleoid the blockage of transcription are but different manifestations of the same activity.

Base Sequence↗
PubMed1985

Isolation and characterization of the Streptomyces cattleya temperate phage TG1.

A temperate actinophage, TG1, was isolated from soil by growth on Streptomyces cattleya and has been shown to be potentially useful for the cloning of DNA in this organism and other streptomycetes. It forms stable lysogens by integration at a unique site on the chromosome. The phage genome consists of 41 kb of double-stranded DNA with cohesive ends. It has unique sites for ClaI, NdeI, PstI, SmaI, and XbaI. The PstI site has been shown to be in a dispensable region of the phage genome. Deletions (2 kb in length) were obtained which retain this site and should be useful for the cloning of DNA.

Attachment Sites, Microbiological↗
PubMed1984

Bacteriophage T4 gol site: sequence analysis and effects of the site on plasmid transformation.

The Escherichia coli lit gene product is required for the multiplication of bacteriophage T4 at temperatures below 34 degrees C. After infection of a lit mutant host, early gene product synthesis is normal, as is T4 DNA replication; however, the late gene products never appear, and early gene product synthesis eventually ceases. Consequently, at late times, there is no protein synthesis of any kind (W. Cooley, K. Sirotkin, R. Green, and L. Snyder, J. Bacteriol. 140:83-91, 1979; W. Champness and L. Snyder, J. Mol. Biol. 155:395-407, 1982), and no phage are produced. We have isolated T4 mutants which can multiply in lit mutant hosts. The responsible T4 mutations (called gol mutations) completely overcome the block to T4 gene expression (Cooley et al., J. Bacteriol. 140:83-91). We have proposed that gol mutations alter a cis-acting regulatory site on T4 DNA rather than a diffusible gene product and that the wild-type form of the gol site (gol+) somehow interferes with gene expression late in infection (Champness and Snyder, J. Mol. Biol. 155:395-409). In this communication, we report the sequence of the gol region of the T4 genome from five different gol mutants. The gol mutations are all single-base-pair transitions within 40 base pairs of DNA. Therefore, the gol site is at least 40 base pairs long. The sequence data confirm that the gol phenotype is not due to an altered protein. We also report that the gol+ site in plasmids prevents transformation of Lit- but not Lit+ E. coli. Thus, the gol site is at least partially active in the absence of the T4 genome.

Base Sequence↗
PubMed1984

Genetic and physiological studies of an Escherichia coli locus that restricts polynucleotide kinase- and RNA ligase-deficient mutants of bacteriophage T4.

The RNA ligase and polynucleotide kinase of bacteriophage T4 are nonessential enzymes in most laboratory Escherichia coli strains. However, T4 mutants which do not induce the enzymes are severely restricted in E. coli CTr5X, a strain derived from a clinical E. coli isolate. We have mapped the restricting locus in E. coli CTr5X and have transduced it into other E. coli strains. The restrictive locus seems to be a gene, or genes, unique to CTr5X or to be an altered form of a nonessential gene, since deleting the locus seems to cause loss of the phenotypes. In addition to restricting RNA ligase- and polynucleotide kinase-deficient T4, the locus also restricts bacteriophages lambda and T4 with cytosine DNA. When lambda or T4 with cytosine DNA infect strains with the prr locus, the phage DNA is injected, but phage genes are not expressed and the host cells survive. These phenotypes are unlike anything yet described for a phage-host interaction.

Coliphages↗
PubMed1984

Progesterone-like effects of estradiol on reproductive behavior and hypothalamic progestin receptors in the female rat.

During the rat estrous cycle, estradiol (E2) and progesterone (P) synergize to activate reproductive behavior. However, receptivity and proceptivity can be elicited by E2 alone in ovariectomized (OVX) animals, particularly when E2 doses are high. The purpose of this study was to determine the neuroendocrine mechanism by which E2 elicits P-dependent reproductive behavior. Adult OVX females received estrogen treatment for 72 h, which consisted of 5 mm Silastic capsules containing 100% E2 or 10% E2, or of 3 injections of estradiol benzoate (EB; 20 micrograms daily). At 72 h, animals were sacrificed for nuclear progestin receptor (NPR) measurements, while others were tested for reproductive behavior. The remaining animals received 1-mg injections of E2, P, moxestrol (Mox) or oil, and either were sacrificed 2 h later for NPR measurements or were tested 4 h later for reproductive behavior. A subset of the animals receiving 1 mg E2 received concurrent administration of the protein synthesis inhibitor, anisomycin (ANI; 100 mg/kg). Acute administration of 1 mg of E2 or P significantly elevated proceptivity, receptivity and NPRs in the mediobasal hypothalamus-preoptic area (MBH-POA) and pituitary (PIT) in females primed with 100% E2. An equivalent dose of Mox was without effect. ANI blocked the acute activation of feminine reproductive behavior by 1 mg of E2. In the absence of acute steroid administration, animals primed for 72 h with EB showed higher levels of reproductive behavior than animals primed with 100% E2.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Correlation of muscarinic receptor induction in the ventromedial hypothalamic nucleus with the activation of feminine sexual behavior by estradiol.

The administration of the steroid hormone, 17 beta-estradiol (E2) to ovariectomized rats increases the levels of muscarinic cholinergic receptors by 20-40% in the hypothalamic area. Using microdissection techniques, this increase has been shown to occur in hypothalamic subregions that possess high levels of intracellular receptors for E2. Among these regions is the ventromedial nucleus (VMN), which may be the principal target site in the rat brain for the activation of feminine sexual behavior by E2. In this study, we have further characterized the increase in muscarinic binding within the VMN after E2 and show that it satisfies many of the criteria for an important regulatory mechanism by which E2 could activate sexual behavior: (1) Using quantitative receptor autoradiography, it was determined that the increase in [3H]-QNB binding in the VMN after E2 results from an increase in the number of receptors rather than a change in the affinity of the binding. (2) The increase in VMN muscarinic receptors as determined by the binding of [3H]-QNB to microdissected homogenates occurred by 18-24 h after exposure to E2, the earliest time at which sexual behavior is facilitated. (3) Increased muscarinic receptors did not occur in the VMN of male rats, which show little activation of feminine sexual behavior after E2 exposure. (4) Muscarinic receptors in the VMN of female rats are induced at 24 h by a 6 h exposure to E2, the minimum length of time that is sufficient for E2 to activate sexual behavior. A 4 h exposure to E2 is insufficient to activate both sexual behavior and induce muscarinic receptors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1983

Serotonergic and noradrenergic receptors in the rat brain: modulation by chronic exposure to ovarian hormones.

Noradrenergic (alpha 1 and beta) and serotonergic (5HT1 and 5HT2) receptors were assayed in the brains of ovariectomized female rats treated for 2 weeks with estrogen, progesterone or a combination of both hormones. Estrogen treatment resulted in a decrease in the number of 5HT1 and beta adrenergic receptors, with a concomitant increase in 5HT2 receptors. Progesterone alone caused a smaller increase in 5HT2 receptors, a similar decrease in 5HT1 and had no significant effect on noradrenergic receptors. When given with estrogen, progesterone blocked the estrogen effect on 5HT2 receptors but did not inhibit the estrogen-mediated decrease in 5HT1 and beta adrenergic receptors. alpha 1 adrenergic receptors were not affected by any of the hormone treatment paradigms. beta adrenergic and 5HT2 receptors are often implicated in antidepressant action, and the modulation of these two receptor types by ovarian hormones might be relevant to hormone-linked affective changes such as premenstrual tension and post-partum depression.

Animals↗
PubMed1982

Competition by estrogens for catecholamine receptor binding in vitro.

We have examined the ability of various steroids to compete for high-affinity binding of 3H-labeled ligands to catecholamine receptors in membranes prepared from rat cerebral cortex, striatum, and anterior pituitary. Ligands employed were: [3H]WB4101, [3H]prazosin, [3H]yohimbine, and [3H]clonidine (alpha-noradrenergic); [3H]dihydroalprenolol (beta-noradrenergic); [3H]spiperone and [3H]ADTN (dopaminergic). Only the 17 beta estrogens were effective and only binding of [3H]spiperone and [3H]ADTN in striatum and [3H]WB4101 and [3H]prazosin in cerebral cortex was reduced. Thus putative dopaminergic and alpha 1-noradrenergic sites alone appear to recognize estrogens. A slight competitive effect on [3H]spiperone binding to anterior pituitary membranes was also observed. Among the 17 beta estrogens tested, the most effective in all cases was the catechol estrogen 2-hydroxyestradiol (2-OHE2). The ability of 2-OHE2 (IC50 = 20-30 micro M) to inhibit ligand binding to alpha 1 receptors was comparable to that of norepinephrine (IC50 = 10-20 micro M), whereas for dopamine receptors in striatum and pituitary 2-OHE2 was an order of magnitude less effective than dopamine (IC50 = 12 micro M) in reducing binding of 3H ligands. Estradiol-17 beta and 2-hydroxyestrone were also able to inhibit binding, but the order of steroid potency was different for alpha 1 and dopaminergic receptors. Progesterone, testosterone, and corticosterone were without effect in all cases. These results show that there is specificity of steroid interactions with catecholamine receptors in the brain, both in terms of steroid structure and receptor type. The possible relevance of these interactions to neuroendocrine function is discussed.

Animals↗
PubMed1980

Shutoff of lambda gene expression by bacteriophage T4: role of the T4 alc gene.

Bacteriophage T4 normally contains 5-hydroxymethylcytosine instead of cytosine in its DNA. Multiple mutants of T4 which synthesize DNA with cytosine do not transcribe their late genes due to the action of the T4 alc gene (Snyder et al., Proc. Natl. Acad. Sci. U.S.A. 73:3098--3102, 1976), which is also responsible for unfolding the host nucleoid after T4 infection (Sirotkin et al., Nature [London] 265:28--32, 1977; Tigges et al., J. Virol. 24:775--785, 1977). It seems reasonable that T4 alc function plays a role in shutting off host transcription, and the observation that some of the RNA made after infection with a T4 alc mutant hybridizes to Escherichia coli DNA (Sirotkin et al., Nature [London] 265:28--32, 1977; Tigges et al., J. Virol. 24:775--785, 1977) supports this hypothesis. Although it is likely that the roles of the alc function in the blocking of some types of transcription and in the unfolding of the host nucleoid are related, it is not known how these effects are achieved or, in fact, whether all types of transcription are affected equally by the alc function. In an attempt to answer these questions, we studied the effect of T4 alc function on bacteriophage lambda transcription and on the structure of intracellular lambda DNA. We found that the alc function is responsible for the shutoff of lambda late transcription but probably not for the shutoff of lambda early transcription. We also found that alc does not block lambda transcription by directly removing the supercoils from circular lambda DNA via either a nicking or topoisomerase activity. Furthermore, we conclude that T4 infection also prevents the translation of non-T4 mRNA because late lambda mRNA's were made after superinfection by a T4 alcs mutant and were of normal length but were not translated into lambda late proteins.

5-Methylcytosine↗
PubMed1978

Isolation of a bacterial host selective for bacteriophage T4 containing cytosine in its DNA.

An Escherichia coli B strain, B834 galU56, has been isolated which supports growth of bacteriophage T4 with cytosine in its DNA while restricting growth of T4 with hydroxymethylcytosine. This host is partially deficient in uridine diphosphoglucose as determined by the ability of DNA isolated from T4 grown on it to accept glucose in an in vitro assay. In this mutant an intact rgl restriction system recognizes unglucosylated hydroxymethylcytosine residues in phage DNA, while the absence of a functional rB restriction function prevents degradation of unmodified DNA containing cytosine.

Coliphages↗
PubMed1977

Postmortem pink teeth.

A series of cases is reported in which pink teeth were observed during the postmortem period. Most cases were associated with decomposition in a moist environment. Experimental procedures led to the extraction of pink material from dentin and demonstration that hemoglobin and serum proteins were present. The pink-tooth phenomenon was duplicated in human teeth by instilling into the pulp chambers whole blood and blood with the red cells hemolyzed. The change was manifested in teeth of dogs after freezing, heating, and decomposition in a moist environment. The authors postulate that pink teeth occur as a result of breakdown of red blood cells in the pulp chamber of the tooth and diffusion of hemoglobin and other serum proteins into the dentin via the dential tubules. Histochemical studies show that the brown or gray material in some teeth subjected to postmortem aging is probably hemoglobin and serum proteins. Factors of age, vascularity of the pulp chamber, and postmortem conditions are discussed in relation to the postmortem development of pink teeth.

Adult↗