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R A Young

Publications and source records attributed to R A Young.

At least 19 recordsLinked to original sources
PubMed1979

Transcription termination in the Escherichia coli ribosomal RNA operon rrnC.

The distal portion of the Escherichia coli ribosomal RNA operon rrnC has been analyzed to determine (a) the transcription boundaries of an operon containing rRNA genes distal to rDNA, and (b) the sequence elements which dictate efficient termination of RNA synthesis. I have sequenced a 505-base pair DNA restriction fragment containing the genes which specify 5 S rRNA, TRNA1Asp, and tRNATrp, in that order. Segments of a spacer DNA separate these genes: the 5 S-tRNA1Asp and tRNA1Asp-tRNATrp spacers are 52 and 8 base pairs long, respectively. In vitro transcription of restriction endonuclease fragments containing the distal portion of rrnC indicates that termination of transcription occurs at a locus beginning 28 base pairs after the tRNATrp gene. The rrnC terminator shares several features with many other known sites of transcription termination in E. coli.

Base Sequence↗
PubMed1979

Sequence of the 16 S-23 s spacer region in two ribosomal RNA operons of Escherichia coli.

The transducing phages lambdadaroE and lambdadilv5, which carry the Escherichia coli ribosomal RNA operons rrnD and rrnX, respectively, have been mapped with the restriction endonucleases BamHI, EcoRI, HindIII, and Sma I. Using hybridization techniques, we have located the ribosomal RNA genes on these phage DNAs. The DNA sequence of the 437-base-pair 16 S-23 S ribosomal RNA intergenic spacer in the two rRNA operons rrnD and rrnX has been determined. The nucleotides examined exhibit only one base pair change between rrnD and rrnX. Both spacer regions contain the genes for tRNA1Ile and tRNA1BAla; the gene sequences are identical with the previously deduced tRNA sequences and are clustered within the first 60% of the spacer DNA. The most striking feature of the 16 S-23 S intergenic region in these two operons is the disparity in G-C content between the tRNA gene sequences (60% G-C) and the remaining spacer DNA (37% G-C). Spacer sequences are known to be involved in the processing of the ribosomal RNA transcript by RNase III and RNase P. In addition, we report the sequence of the first 108 base pairs of the 23 S rRNA gene.

Base Sequence↗
PubMed1979

Tandem promoters direct E. coli ribosomal RNA synthesis.

To determine the special feature of ribosomal RNA promoters that might account for the highly efficient and regulated synthesis of rRNA in E. coli, we have analyzed the beginnings of two ribosomal RNA operons, rrnD and rrnX. DNA sequences for 425 bp preceding those specifying mature 16s rRNA are reported. In vitro transcription of restriction endonuclease fragments containing this region from either operon reveals the presence of two promoters about 110 nucleotides apart; they are denoted P1 and P2. RNA synthesis from P1 is initiated with GTP at position -284 (relative to 16s sequences) in rrnD and with ATP at position -285 in rrnX. At P2, the RNA starts with CTP primarily at position-176 in both operons. The DNA sequences of the two operons are identical for 231 bp preceding the 16s rDNA (including a substantial region around P2); they then diverge almost completely, except for a notable 18 bp homology just preceding the transcription start site for P1. Certain sequences implicated in the recognition of promoters by E. coli RNA polymerase are clearly identifiable in both P1 and P2; other features include an extended region preceding P1 which is strikingly rich in AT base pairs. Possible mechansims by which these tandem promoters contribute to the high frequency of rRNA transcription and to the differential expression of the E. coli rrn operons are discussed.

Base Sequence↗
PubMed1979

Transposition of the Escherichia coli insertion element gamma generates a five-base-pair repeat.

We have determined DNA sequences surrounding the termini of the Escherichia coli insertion element gamma delta, both at its normal locus on the F (fertility) factor and at three different sites of insertion into the plasmid pBR322. After transposition, a five-base-pair pBR322 sequence is duplicated and appears in direct orientation adjacent to each end of the element. No such duplication flanks the ends of gamma delta in F, and there is no apparent homology between the sequences surrounding gamma delta in F and the five-base-pair duplications generated by insertion. These findings suggest that the duplications are not essential for transposition and that they do not act to direct gamma delta to a homologous site in the target chromosome. In addition, we find that the 35-base-pair inverted repeat that comprises the termini of gamma delta is strikingly similar in sequence to the ends of both the ampicillin-resistance transposon Tn3 and a 200-nucleotide-long sequence on the plasmid pSC101 which has been shown to mediate recombination with phage f1 replicative form. Within the terminal region, there is a specific heptanucleotide sequence common to each of the above elements and to bacteriophage Mu, all of which generate five-base-pair repeats upon insertion.

Base Sequence↗
PubMed1979

Seasonal variation and the influence of body temperature on plasma concentrations and binding of thyroxine and triiodothyronine in the woodchuck.

Woodchuck plasma was collected during four seasons of the year and assayed for total and dialyzable (free) T4 and T3 and for rT3. Plasma concentrations of total and free T4 and T3 were higher in the spring (T4, 5.4 +/- 0.6 microgram/dl; free T4, 3.0 +/- 0.4 ng/dl; T3, 202 +/- 22 ng/dl; free T3, 0.51 +/- 0.04 ng/dl) and lower in the prehibernatory fattening period in summer (T4, 2.3 +/- 1.0 microgram/dl; free T4, 1.2 +/- 0.5 ng/dl; T3, 45 +/- 27 ng/dl; free T3, 0.16 +/- 0.10 ng/dl) and fall (T4, 3.2 +/- 1.0 microgram/dl; free T4, 1.3 +/- 0.2 ng/dl; T3, 130 +/- 12 ng/dl; free T3, 0.25 +/- 0.02 ng/dl). In spite of the extremely high concentrations of T3 in the winter (437 +/- 32 ng/dl), free T3 concentrations (0.034 +/- 0.003 ng/dl), when measured at the appropriate temperature for hibernation, were significantly lower than those found at other seasons of the year. Plasma binding of T3 was lower during the summer and increased again to approximately double the spring value during the winter. rT3 was at or below the sensitivity of the method (6 ng/dl) at all seasons. It is suggested that the wide seasonal variations in thyroid hormone concentrations and altered plasma protein binding may represent important adaptations influencing the metabolic rate and the process of hibernation in the woodchuck.

Animals↗
PubMed1979

The woodchuck, Marmota monax, as a laboratory animal.

The woodchuck or groundhog (Marmota monax) has been used as a biomedical model for studies of obesity and energy balance, endocrine and metabolic function, central nervous system control mechanisms and cardiovascular, cerebrovascular and neoplastic disease. Methods of care of a woodchuck colony, techniques for handling, restraint, anesthesia, blood sampling and breeding were developed.

Animal Husbandry↗
PubMed1978

Seasonal ultrastructural variations in pinealocytes of the woodchuck, Marmota monax.

The ultrastructure of the pinealocyte in the woodchuck, Marmota monax, was studied during the four seasons of the year. Fall cells have a fairly uniform cytoplasmic density, organelles consistent with synthetic and/or secretory activity and rather extensive pericapillary and intercellular spaces. Many winter pinealocytes are nearly devoid of ribosomes and granular endoplasmic reticulum but contain lipid droplets associated with mitochondria. Pericapillary and intercellular spaces are minimal. Spring glands have the greatest variation in cytoplasmic density with intercellular and pericapillary spaces similar to that seen in fall glands. Cells containing electron dense cytoplasm have Golgi zone associated, secretory granules, free ribosomes, short sections of granular endoplasmic reticulum and dense bodies. Cells with a more electron lucent cytoplasm are similar to the most frequently observed summer pinealocytes which have numerous Golgi zones but few associated secretory granules. Microtubules are prominent in the cytoplasm of these cells, the plasma membranes are smooth and intercellular and pericapillary spaces are minimal. A yearly rhythm or cyclic activity of the pinealocyte is suggested.

Animals↗
PubMed1978

The parathyroid gland of the woodchuck (Marmota monax): a study of seasonal variations in the chief cells.

The ultrastructure of the parathyroid chief cell in the woodchuck, Marmota monax, was studied during the four seasons of the year. Spring chief cells have stacks of granular endoplasmic reticulum, prominent multiple Golgi zones and many clumped mitochondria. Summer cells resemble those seen in the spring but the mitochondria are associated with stacks of granular endoplasmic reticulum. Multiple areas of stacked granular endoplasmic reticulum characterize the fall chief cells. Their Golgi zones are large and are associated with many dense core secretory granules. Lipoid vacuoles are frequently noted. Winter chief cells have secretory granules and phagolysosomes (dense bodies). Some of these cells contain stacked arrays of granular endoplasmic reticulum associated with mitochondria, others have only short segments. The above morphological findings are discussed in relation to those in other hibernators, the parafollicular (C) cell, and to the cyclic seasonal activities of the woodchuck.

Animals↗
PubMed1978

Complementary sequences 1700 nucleotides apart form a ribonuclease III cleavage site in Escherichia coli ribosomal precursor RNA.

The nucleotide sequence of Escherichia coli DNA at both ends of the gene for 16S rRNA has been determined for two rRNA operons, rrnD and rrnX. The 400 nucleotides we have examined exhibit only one base change between rrnD and rrnX. Within the 160 nucleotides that precede mature 16S rRNA sequences are cleavage sites for several E. coli endonucleases, including RNase III. A 240-nucleotide segment encompassing the 16S 3' end contains another RNase III site and the point of presumed RNase P scission at the 5' end of tRNA1Ile, the first tRNA appearing in the 16-23S spacer region of rrnD and rrnX. Most importantly, the DNA sequences predict that regions flanking the 16S gene in the rRNA primary transcript extensively base pair to form a double-helical structure whose hairpin loop includes the entire mature 16S molecule; within this structure is a 26-base-pair stem containing the two sequences at which RNase III action generates the 5' and 3' ends of a previously characterized precursor to 16S rRNA. Although our proposed secondary structure for this RNase III site is superficially dissimilar to previously described cleavage sites in the T7 early mRNA precursor, certain common features may constitute signals for RNase III recognition. The suggestion that distant portions of an RNA molecule can form a secondary structure within which specific endonucleolytic cleavages occur may have mechanistic implications for the joining of noncontiguous portions of gene sequences evident in several eukaryotic mRNAs.

Base Sequence↗
PubMed1977

Host factor for coliphage Q beta RNA replication: presence in procaryotes and association with the 30S ribosomal subunit in Escherichia coli.

The Host Factor required for in vitro coliphage Q beta RNA replication, a heat-stable RNA binding protein present in uninfected Escherichia coli, has been detected by both immunological and functional tests in Acinetobacter calcoaceticus, Klebsiella pneumoniae, Pseudomonas aeruginosa and Pseudomonas putida. It was not detectable by these criteria in Bacillus stearothermophilus, Bacillus subtilis, Caulobacter crescentus, Micrococcus lysodeikticus, Rhodopseudomonas capsulata or Saccharomyces cerevisiae. In Escherichia coli the Host Factor protein has been shown to be associated with ribosomes. It is demonstrated here that this association is specific for the 30S ribosomal subunit.

Acinetobacter↗
PubMed1977

The thyroid gland of the woodchuck, Marmota monax: a morphological study of seasonal variations in the follicular cells.

The morphology of the thyroid gland of the woodchuck, Marmota monax, was studied during the four seasons of the year. In the spring the thyroid is extremely heterogenous in appearance. Some follicular cells appear quite active. They contain a well defined Golgi apparatus, abundant large colloid droplets and pseudopodia but few, if any, apical vesicles. Other less active cells have poorly defined rough surfaced endoplasmic reticulum and lack a well developed Golgi apparatus. They do not contain apical vesicles or colloid droplets. Summer thyroids have uniformly small follicles which are lined by high cuboidal cells containing numerous mitochondria, apical vesicles, abundant rough surfaced endoplasmic reticulum, and lipid droplets but few colloid droplets. There is extensive lateral and basal infolding of the cytoplasmic membranes in these cells. In the fall and winter the follicles are larger than in the summer and contain more colloid. Numerous heterogeneous dense bodies appear in the cytoplasm of the follicular cells in the fall and increase in number in the winter when there is an obvious sparsity of such glycoprotein synthetic organelles as Golgi apparatus and rough surfaced endoplasmic reticulum. These morphologic changes are compared with previous studies of thyroid structure and function in other animals and are correlated with the seasonal physiologic activities of the woodchuck.

Animals↗
PubMed1977

Seasonal variation in the morphology of thyroid parafollicular (C) cells in the woodchuck, Marmota monax: a light and electron microscopic study.

The morphology of parafollicular (C) cells in the thyroid gland of the woodchuck, Marmota monax, was studied during the four seasons of the year. The spring C cells are characterized by a large Golgi zone, rough-surfaced endoplasmic reticulum, free ribosomes and relatively few dense granules. In the summer these cells appear to be larger and many are packed with dense granules. Fall cells exhibit morphological characteristics suggestive of intense synthetic activity, having rough-surfaced endoplasmic reticulum in whorls or parallel arrangement, a large Golgi apparatus and few, to many granules, some of which are larger than those seen in the spring and summer C cells. In the winter, most of the C cells are packed with granules. The Golgi zone, when observed, is small and the rough-surfaced endoplasmic reticulum is sparse. Many cell profiles exhibit apparent granule dissolution. These cyclic morphological findings are discussed along with previous studies of other hibernators and are correlated with the seasonal activities of the woodchuck.

Animals↗
PubMed1977

Leiomyosarcomatosis of probable uterine origin with long survival--a case report.

A case of leiomyosarcomatosis is presented. Over a period of 15 years the patient underwent seven operations to remove eleven tumours, the largest as big as a football, before dying of widespread metastases: between operations the patient was remarkably well. The disease was almost certainly of uterine origin from apparently benign fibroids, and photomicrographs are provided as supportive evidence. The various modes of presentation of abdominal leiomyosarcoma--mass, pain, obstruction, fistula, anaemia--are illustrated in the case report. The value of repeated palliative surgery in such cases is emphasized.

Female↗
PubMed1976

Function and structure in phage Qbeta RNA replicase. Association of EF-Tu-Ts with the other enzyme subunits.

Qbeta replicase is a complex of four nonidentical subunits readily dissociable into two subcomplexes: 30 S ribosomal protein S1 and the phage-coded polypeptide (Subunits I + II) and protein synthesis elongation factors EF-Tu and EF-Ts (Subunits III + IV). The affinity of the two subcomplexes for one another increases with increasing ionic strength. The enzyme is capable of initiation of RNA synthesis with synthetic templates only when in the low ionic strength conformation. Elongation of initiated polynucleotide chains is not affectedby ionic strength. Addition of Qbeta RNA to the enzyme also alters its quaternary structure: the EF-Tu-Ts cannot be covalently attached to the other enzyme subunits with bifunctional cross-linking reagents in the presence of RNA. This conformational change is not influenced by ionic strength. The addition of Qbeta RNA to the enzyme, does not result in the release of EF-Tu-Ts from the other enzyme subunits: whereas free EF-Tu-Ts binds GDP independently of salt concentration, this binding by Qbeta replicase is sensitive to high ionic strength and remains so in the presence of Qbeta RNA. Furthermore, RNA does not allow the release of EF-Ts from EF-Tu by GTP as measured by sensitivity of EF-Ts activity to N-ethylmaleimide.

Binding Sites↗