Poly(d(A—T)) dependent RNA polymerase activity after treatment of nuclei with micrococcus nuclease

• 1.1. Rat liver nuclei were incubated with or without 20 units micrococcus nuclease (EC3.1.4.7)/mg nuclear DNA.
• 2.2. The soluble poly(d(A—T)) dependent RNA polymerases were reduced in activity to 15–20% that of the controls after treatment with micrococcus nuclease.
• 3.3. RNA polymerases I plus III activities were completely, RNA polymerase II activity partially reversible on removal of the DNA released into the soluble fraction by treatment of nuclei with micrococcus nuclease.
• 4.4. Inhibitory constants obtained with the solubilized DNA were 17.1 μM and 20.7 μM nucleotide-DNA for RNA polymerases I plus III and RNA polymerase II, respectively. The corresponding inhibitory constants obtained with native salmon DNA were 23.0 μM and 34.4 μM nucleotide-DNA.

     

Comparison of nuclear DNA-dependent rna polymerases from mouse tumors and tissues of normal swiss mice

• 1.1. Five major forms of nuclear DNA-dependent RNA polymerases from seven transplantable murine tumors and from five tissues from normal, adult, male Swiss mice (Mus musculus) were separated chromatographically on DEAE-Sephadex A-25. Forms Ia and Ib were insensitive to α-amanitin, whereas forms IIa and IIb were highly sensitive and form III was slightly sensitive.
• 2.2. The polymerases from all tumors or ascites tumors only were compared statistically with those from the normal tissues in regard to elution patterns, specific activities, activities per mg of nuclear DNA. degrees of purification and yields. Similarly, individual tumors were compared with homologous normal tissues. All parameters were characterized by relatively large variance.
• 3.3. Activities of all RNA polymerases per mg DNA were higher in all tumors compared with normal tissues. The order of statistical significance of these differences was lib > III > IIa > Ib > Ia.
• 4.4. Activities per mg DNA of all RNA polymerases from 6C3HED and L1210 tumors, with the exception of L1210 tumor enzyme III, were significantly greater than those from spleen, but only the activities of Lewis lung tumor enzyme IIb and of Taper hepatoma enzymes Ia and III were higher than those from the homologous tissues, lung and liver, respectively.

     

A comparative study of nuclear and chloroplast DNA dependent RNA polymerases from wheat leaves

• 1.1. Three DNA dependent RNA polymerases have been purified from chromatin and chloroplast fractions of wheat leaves.
• 2.2. The purified enzymes were completely dependent on exogenous DNA after purification by glycerol gradient, DEAE-Sephadex and phosphocellulose chromatography.
• 3.3. The nuclear enzymes, I and II, showed a strong preference for denatured nuclear DNA, whereas the chloroplast enzyme preferred denatured chloroplast DNA.
• 4.4. The three enzymes require either Mg²⁺ or Mn²⁺ for activity.
• 5.5. α-amanitin specifically inhibited RNA polymerase II but has no effect on polymerase I and chloroplast polymerase.
• 6.6. Enzyme I is most active at very low ionic strength (0.10 mM KC1), whereas enzyme II and chloroplast enzyme show maximum activity at 150mM and 50 mM KC1 respectively.

     

Age-related changes in total dna and rna and incorporation of uridine and thymidine in rat liver,kidney and spleen

• 1.1. Total content of DNA and RNA in liver, kidney and spleen were measured in young and aged rats. At the same time the incorporation of [¹⁴C]thymidine, a DNA precursor, and [³H]uridine, an RNA precursor, were also determined.
• 2.2. Changes in total organ DNA and RNA correlated with sexual maturation as did incorporation of precursors.
• 3.3. Young animals have more DNA per organ relative to RNA. with kidney and spleen DNA showing a decrease between maturity and senescence.
• 4.4. However, liver RNA increases with age. a change probably due to decreased catabolism of RNA since [³H]uridine uptake decreases.
• 5.5. Liver polyploid differentiation, and [¹⁴C]thymidine and [³H]uridine uptake, are correlated.
• 6.6. In kidney, incorporation of [³H]uridine is inversely related to [¹⁴C]thymidine incorporation.

     

A study on the rna polymerase activities in bovine thyroid nuclei isolated in isotonic sucrose,hypertonic sucrose or in anhydrous glycerol media: Influence of the isolation procedure on the electrophoretic pattern of acid soluble nuclear proteins

• 1.1. After differential pelleting of bovine thyroid bound RNA polymerase II was the more enriched enzyme activity in the nuclear fraction, and coincided best with the DNA profile.
• 2.2. The RNA polymerase I + III activity was compared in nuclear fractions isolated either in 0.25 M sucrose (wet tissue) or in anhydrous glycerol (lyophilized tissue) or in 2.4 M sucrose (lyophilized tissue).
• 3.3. Although the nuclei were more resistant to the isolation porcedure in glycerol, more proteins were extracted by that procedure than during the isolation in 2.4 M sucrose.
• 4.4. With the 2.4 M sucrose method a twofold enrichment of RNA polymerase I + III activity in respect to DNA occurred in the nuclei pointing to an exclusive localization of these activities within the nucleus.
• 5.5. Using the same isolation procedure the different classes of histones were better resolved upon polyacrylamide gelelectrophoresis.

     

Lipid changes in blood serum and tissues of the Egyptian Cobra “Naja haje haje” during the hibernation cycle

• 1.Total lipids, free fatty acids, triglycerides, phospholipids and total cholesterol in blood serum, liver, brain, cardiac and skeletal muscles of Naja haje haje were determined during the different phases of the hibernation cycle.
• 2.A sharp decrease in the level of total lipids of blood serum and all tissues occurred during hibernation. Upon arousal, lipogenesis is commonly restored.
• 3.Elevated concentrations of serum free fatty acids predominated in pre-hibernation and hibernation periods, while the tissues recorded highly significant declines during hibernation.
• 4.Occurrence of marked decreases in triglycerides contents of serum and tissues except the cardiac muscles in the hibernation and arousal phases.
• 5.Sharp increases in the phospholipid contents of blood and the selected tissues were recorded during hibernation. The level declined in both liver and cardiac muscles in arousing animals.
• 6.Total cholesterol level was lowered in blood during hibernation. The cardiac muscles showed a highly significant decrease while liver, brain and skeletal muscles showed elevations in the same phase.

     

Supply of polyenoic fatty acids to the mammalian brain: The ease of conversion of the short-chain essential fatty acids to their longer chain polyunsaturated metabolites in liver,brain, placenta and blood

The metabolism of linoleic and linolenic acids to the longer polyunsaturated fatty acids of mammalian brain is discussed. Differences in metabolic activity are considered between tissues, between species, and during different stages of development. Available evidence suggests that:

• 1.1. The sites of metabolism are confined mainly to liver and the brain itself.
• 2.2. The similar metabolic pathways are subject to a complex control which is most effective at the first step in the sequence.
• 3.3. During early development there are reciprocal changes in the metabolic capacity of brain and liver.
• 4.4. Metabolic activity varies between species and may be absent in obligate carnivores.

     

Hybridization properties of non-polyadenylated oligo(U)-containing RNA from germinating wheat embryos

• 1.1. Total cytoplasmic RNA of germinating wheat embryos was fractionated by affinity chromatography and separated into non-polyadenylated oligo(U)-containing RNA (A(−)U(+)RNA) and polyadenylated oligo(U)-lacking RNA (A(+)U(−)RNA).
• 2.2. The reassociation kinetics of ³²P-labelled complementary DNA (cDNA) reverse-transcribed from A(−)U(+)RNA shows that this RNA fraction is transcribed from unique DNA sequences of the genome similarly as typical mRNA.
• 3.3. Cross hybridization experiments show no significant sequence homology between the two RNA fractions. Therefore it is concluded that non-polyadenylated oligo(U)-containing RNA of wheat embryo may represent a discrete class of mRNA.

     

Relative bioavailability and DNA adduct formation of benzo[a]pyrene and metabolites in the diet of the winter flounder

• 1.1. Radiolabeled metabolites of the carcinogenic polycyclic aromatic hydrocarbon benzo[a]pyrene (BaP) were shown to be absorbed through the diet of the winter flounder, Pseudo pleuronectes americanus.
• 2.2. Oral bioavailability of a mixture of naturally produced metabolites was significantly less than that of the parent BaP.
• 3.3. Oral bioavailability of a pure metabolite, BaP-7,8-dihydrodiol (7,8-D) was found to be similar to that of BaP.
• 4.4. Both metabolites and BaP formed DNA adducts in liver.

     

Comparison of U-small nuclear RNA levels in tissues of the silkmoth,Bombyx mori

• 1.1. Tissue-specific abundance of the capped small RNAs in the silkmoth Bombyx mori was compared using preparative immunoprecipitation with anti-trimethylguanosine antibody.
• 2.2. The yields of total capped small RNAs from larval posterior silk gland, 1. early, 2. late in the fifth-instar, and 3. immortal ovarian-derived cells in culture, were determined to be 187, 50 and 218 ng, respectively, per mg of total cellular RNA.
• 3.3. Separation of immunoprecipitated RNAs by polycrylamide gel electrophoresis, followed by densitometric analysis of the bands, allowed the quantitation of individual capped molecules.
• 4.4. This analysis revealed tissue-specific patterns.
• 5.5.|The data indicate that the total abundance of capped small RNAs in Bombyx is highest in rapidly-dividing cells.

     

Alcohol dehydrogenase ontogeny and liver maturation in the leopard danio,brachydanio nigrofasciatus

• 1.1. The tissue specificity and ontogeny of alcohol dehydrogenase (ADH) are reported for the leopard danio, Brachydanio nigrofasciatus.
• 2.2. Of the seven adult tissues assayed (eye, brain, kidney, liver, ovary, skeletal muscle and stomach), only liver extracts showed ADH activity.
• 3.3. The activation of the Adh locus is correlated with the first functioning of liver. It is suggested that the state of liver cell differentiation may be the stimulus necessary for Adh locus expression.

     

Lactate dehydrogenase isozymes of the leopard danio,Brachydanio nigrofasciatus: their characterization and ontogeny

• 1.1. The tissue specific patterns and ontogeny of lactate dehydrogenase (LDH) are reported.
• 2.2. While all tissues (eye, brain, heart, intestine, liver, ovary and skeletal muscle) show isozymes of A and B subunit composition, only liver extracts possess isozymes resulting from C subunit synthesis.
• 3.3. The A₄ homopolymer appears simultaneously with initial muscle contractility and is correlated with the physiological function of muscular contraction.
• 4.4. The activation of the Ldh-C locus is correlated with the first functioning of liver. It is suggested that the state of differentiation of liver cells may be the stimulus required for C locus expression.

     

Effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail

• 1.1. The effects of niacin deficiency on the relative turnover rates of proteins in various tissues of Japanese quail were investigated.
• 2.2. The level of liver NAD was not affected by niacin deficiency whereas the level of pectoral muscle NAD was markedly reduced.
• 3.3. In all dietary treatments the liver had the highest turnover rates of proteins, heart and brain had intermediate rates, and pectoral muscle had the lowest rates.
• 4.4. Relative turnover rates of proteins in all tissues (particularly pectoral muscle) of the niacin deficient group were significantly higher than those of pair-fed control group, although there were no significant differences in turnover rate between pair-fed control and control groups.
• 5.5. The high turnover rate of proteins in niacin deficiency was primarily attributed to enhanced degradation rate of proteins rather than enhanced synthesis rate of proteins.
• 6.6. Optical density scanning (or densitometric) of water-soluble pectoral muscle proteins separated by isoelectric focusing revealed several additional minor protein bands between major protein bands in the niacin deficient group which were more pronounced in the acidic region of the gel.
• 7.7. These results suggest that proteins with a low pI value in pectoral muscle of the niacin deficient animal are highly sensitive to protein degradation.

     

He partial sequencing of human nonspecific (bone/liver/kidney) alkaline phosphatase gene

• 1.1. A charon 4A human fetal liver genomic library was screened for human nonspecific alkaline phosphatase gene using the cloned human bone cDNA as a hybridization probe.
• 2.2. A clone 2.2 Kb DNA was sequenced and found to contain a piece of sequences encoding the 4–44th amino acids of NH; terminus.
• 3.3. The other cloned 1.6Kb DNA contains two segments of sequences each corresponding to two separate regions of the cDNA for alkaline phosphatase. The first segment of the DNA codes for the 83–141st amino acids whereas the second for 141–199th.

     

Purine nucleoside kinases in mouse tissues and human lymphoid cells in culture

• 1.1. Purine nucleoside kinase activities (adenosine kinase, deoxyadenosine kinase and arabinosyl adenine kinase) in mouse tissues were in the following order: liver > kidney > heart > lung > brain > spleen > intestine.
• 2.2. Ratios of deoxyadenosine or arabinosyl adenine kinase to adenosine kinase were significantly higher (10–200 fold) in human lymphoid cells than in mouse tissues.
• 3.3. Leukocytes from T-cell acute lymphoblastic leukemic patients had 5–20 fold elevated adenosine deaminase as compared to normal T-cells.

     

An electrophoretic comparison of non-histone proteins from rat liver total chromatin and chromatin depleted of 0.35 M NaCl soluble proteins

• 1.1. A procedure of isolation of non-histone proteins from rat liver chromatin in mild conditions provided 3 groups of these proteins, i.e. NHCP1, NHCP2 and NHCP3.
• 2.2. The investigated proteins are devoid of DNA and revealed various influences on RNA synthesis in vitro.
• 3.3. The extraction of rat liver chromatin with 0.35 M NaCl (pH 7.5) removed about 30% of examined proteins. Electrophoretic patterns of 3 groups of non-histone proteins from total chromatin and chromatin depleted of 0.35 M NaCl soluble proteins are compared.

     

Protein tyrosine phosphorylation in normal rat tissues

• 1.1. Exogenous and endogenous tyrosine protein phosphorylation activities were examined in soluble and partieulate fractions from various normal tissues by using poly-[Glu-⁸⁰Na, Tyr²⁰] and a monoclonal antibody specific for phosphotyrosine.
• 2.2. Phosphorylation of the exogenous substrate by the partieulate forms of TPKs was 2- to 10-fold higher than by soluble forms. The activities of partieulate and soluble enzymes decreased in the following order: spleen > (thymus = kidney) > testes ⩾ (pancreas = liver = brain) > heart.
• 3.3. The level of endogenous phosphorylation in the tissues decreased respectively in the following order: thymus > brain ⩾ (pancreas = liver) > spleen > testes > kidney > heart for the partieulate fractions, and spleen > thymus > brain > pancreas ⩾ liver > testes > kidney > heart for the soluble fractions.
• 4.4. A large number of phosphotyrosine-containing proteins were detected. In addition, several phosphotyrosine-containing proteins of similar molecular weight were found in different tissues and fractions.