The size of poly(A)-containing RNAs in Drosophila melanogaster embryos

The size range of poly(A)-containing RNA from Drosophila melanogaster embryos has been estimated by hybridization with ³H-labeled poly(U) and subsequent fractionation on sucrose gradients. The median size of nuclear poly(A)-containing RNA is about 30 S (6000 nucleotides), and the median size of cytoplasmic poly(A)-containing RNA is about 17 S (1800 nucleotides). The relationship of these sizes to messenger RNA needed to code for protein and to the length of DNA contained in a chromomere is discussed.Research grant support was provided by NIH (6M35558; HD-00266) and NSF (GB-30600).     

Series of 4.5S RNAs associated with poly(A)-containing RNAs of rodent cells.

Uninfected mouse kidney cells and mouse leukemia cells L1210 in cluture contained a series of 4.5S RNAs which was structurally identical to the series of 4.5S RNAs associated with genomic RNAs of murine retroviruses and poly(A)-containing RNAs from virus infected cells. Normal rat kidney cells and baby hamster kidney cells in culture also contained a series of 4.5S RNAs. The strucutre of the 4.5S RNAs from mouse, rat and hamster cells were very similar, but not identical. These 4.5S RNAs were not found in cultured cells of other vertebrates, such as human, monkey, cat, mink, rabbit and chicken cells.     

Studies of the cell walls of Schizophyllum commune

Mechanically isolated cell wall materials of eight strains of Schizophyllum commune were studied by chemical and enzymatic procedures. Isolated wall material of each strain was separated by chemical methods into three fractions: A (cold alkali-soluble, , amorphous), B (warm alkali-soluble, amorphous), and C (alkali-insoluble, retaining appearance of hyphal fragments). Chemical tests indicated the presence of chitin in Fraction C and the absence of cellulose, lignin and pectic substances from all fractions. Analyses of acid hydrolysates indicated the presence of glucose in Fractions A, B and C, of hexosamine in Fraction C and the absence of galactose, mannose, 6-deoxyhexoses, xylose and other pentoses from all fractions. Unfractionated material, Fraction A and Fraction B were slightly attacked by commercial cellulase whereas Fraction C was heavily attacked. Commercial chitinase by itself did not attack Fraction C or unfractionated material to a significant extent. In the presence of cellulase, it was active upon Fraction C. Qualitative differences in cell wall composition between strains were not detected; however, quantitative differences were observed in the proportion of Fraction A and Fraction C as well as in the amount of the various breakdown products in certain strains. It is visualized that the cell wall of this fungus consists of a core of chitin covered by or intermeshed with glucose-containing polymers.     

Quantitation and characterisation of poly(A)-containing messenger RNAs from mouse neuroblastoma cells

Comparison of several isolation procedures for neuroblastoma poly(A)-containing mRNAs shows that the highest percentage recovery of undegraded and biologically active messenger RNAs is obtained using proteinase K prior to phenol extraction. The messenger RNAs thus isolated comprise approximately 1.5% of the total ribosomal RNAs and have negligible contamination with 18 and 28 S RNAs. On denaturing polyacrylamide gels they have an average molecular weight of 6.5-10(5) with a range from 2.2-10(5) to 1.53-10(6). The messenger RNAs have an average poly(A) content of 154 nucleotides. They are highly active in wheat germ in vitro protein synthesizing systems, giving as much as 4.3 pmol [35S]methionine incorporation into total protein per mol of mRNA. This is almost as active as a control globin mRNA preparation.     

Nucleotide sequences of 4.5S RNAs associated with poly(A)-containing RNAs of mouse and hamster cells.

The nucleotide sequences of 4.5S RNAs associated with poly-(A)-containing RNAs of mouse and hamster cells were determined. These RNAs have 91 to 94 nucleotides, a high content of G (almost 40%) and no modified nucleoside. The 5'-termini are pppG, but the 3'-termini lack uniformity in the number of uridylate residues. These molecules contain two sets of repeating sequences, and a central purine-rich sequence. There is only one base exchange between mouse and hamster 4.5S RNAs. Possible binding sites of these RNAs to poly(A)-containing RNAs are discussed.     

普通裂褶菌真菌学及实验室研究进展

普通裂褶菌是一种条件致病菌,可侵犯人体多个器官多种组织,导致各种表现形式的疾病,如脑脓肿、脑膜炎、鼻窦炎、肺部真菌球、蜂窝肺、肺结节、甲真菌病等.由于普通裂褶菌感染引起的真菌病病例少见报道,临床医生对其认识不足,再加上某些实验室不具备鉴定条件,所以感染该菌的病例常被漏诊、误诊.现将近年来临床分离的普通裂褶菌株真菌学及实...     

Absence of differences in polysomal RNAs from vegetative monokaryotic and dikaryotic cells of the fungus Schizophyllum commune

Total polysomes, including free and membrane-bound polysomes, were isolated from the monokaryotic and dikaryotic mycelial cell types of the basidiomycete Schizophyllum commune grown under submerged conditions. Sucrose gradient centrifugation showed that these isolated polysomes had a size distribution as expected for polysomes functioning in vivo and thus contained intact mRNA. RNA preparations extracted from the polysomes were used to analyze the mRNA sets in the monokaryon and dikaryon. Saturation hybridization of single-copy DNA with a vast excess of polysomal RNA and cell-free translation of polysomal RNA in a wheat germ system followed by two-dimensional gel electrophoresis of the products did not reveal significant differences between the mRNA sets in both mycelial cell types. Moreover, it was found that the sequence complexities and coding properties of polysomal RNA and total (nuclear and cytoplasmic) RNA from S. commune are not detectably different. From these results and those obtained in previous studies it was concluded that the differentiation between monokaryotic and dikaryotic mycelial cell types as controlled by A and B incompatibility genes involves differential modification of polypeptides during or after mRNA translation.     

The majority of calf muscle cell messenger RNAs contain poly(A)

Previous studies from our laboratory have investigated messenger RNA metabolism in calf muscle cells in tissue culture. The analysis of mRNA was based on its poly(A) content. We have now examined directly the proportion of mRNA which contains poly(A) in these cells. After separation of poly(A)+ -and poly(A) - -RNA on oligo(dT) -cellulos, the two fractions were translated in a reconstituted, heterologous cell-free protein-synthesizing system and the products were compared with those from the translation of total RNA. The great majority of mRNA form either prefusion or postfusion cultures was poly(A)- containing; quantitative determinations show that about 70-90% of the actin mRNA is poly(A)-containing. In order to determine if a large fraction of the calf muscle mRNA can be translated by a heterologous cell-free system, [3H]-POLY(A)+ -RNA was added to reticulocyte lysates and the formation of initiation complexes was followed. These experiments suggest that the bulk of calf muscle cell mRNA would be utilized in such a system and justify the use of cell-free systems to examine the poly(A) content of total mRNA. Thus, differential polyadenylation does not seem to be an important aspect of mRNA metabolism in cultured muscle cells. The previous study of mRNA in these cells, based on poly(A) content, is apparently a valid measure of overall mRNA metabolism.     

Somatic recombination in the common-AB diploid of Schizophyllum commune

Summary A recessive suppressor, su-1, of arg-2 was used to detect somatic recombination in common-AB diploids of S. commune. Recombinants were recovered from dense, fastgrowing sectors on arginine-deficient medium. The majority of spontaneous recombinants (129/154) recovered in this study were apparently haploid. Strains which scored as aneuploid and diploid were also recovered and analyzed. Genetic analysis of spontaneous recombinants indicated that crossing over is rare and that haploidization very likely proceeds via stages of aneuploidy. No increase in the frequency of crossing over was detected in recombinants derived following treatment with UV light. The preliminary results favor a parasexual mechanism of recombination in common-AB diploids.Journal Article No. 5126, Michigan Agricultural Experiment Station. This resarch was supported in part by Contract AT-(11-1) 1301 from the United States Atomic Energy Commission and Grant No. GB-13654 from the National Science Foundation.     

'Cap' structures in maize poly(A)-containing RNA.

Poly(A)-containing RNA was isolated from maize embryos by chromatography on columns of oligo(dT)-cellulose and exhaustively digested with ribonucleases T2, T1, and A. Fractionation of the digests by two-dimensional electrophoresis revealed the presence of three 7-methylguanosine-terminated 'cap structures' of the type m7GpppNp.     

Comparison of cytoplasmic and nuclear poly(A)-containing RNA sequences in Xenopus liver cells.

Poly(A)-containing RNAs from cytoplasm and nuclei of adult Xenopus liver cells are compared. After denaturation of the RNA by dimethysulfoxide the average molecule of nuclear poly(A)-containing RNA has a sedimentation value of 28 S whereas the cytoplasmic poly(A)-containing RNA sediments slightly ahead of 18 S. To compare the complexity of cytoplasmic and nuclear poly(A)-containing RNA, complementary DNA (cDNA) transcribed on either cytoplasmic or nuclear RNA is hybridized to the RNA used as a template. The hybridization kinetics suggest a higher complexity of the nuclear RNA compared to the cytoplasmic fraction. Direct evidence of a higher complexity of nuclear poly(A)-containing RNA is shown by the fact that 30% of the nuclear cDNA fails to hybridize with cytoplasmic poly(A)-containing RNA. An attempt to isolate a specific probe for this nucleus-restricted poly(A)-containing RNA reveals that more than 10(4) different nuclear RNA sequences adjacent to the poly(A) do not get into the cytoplasm. We conclude that a poly(A) on a nuclear RNA does not ensure the transport of the adjacent sequence to the cytoplasm.     

Polymorphism of intracellular isoenzymes of Schizophyllum commune Fr. (Basidiomycetes) in the Donetsk region

The results of a study of the polymorphism of intracellular isoenzymes in cultures of Schizophyllum commune Fr. grown in the Donetsk region are presented here. The electrophoretic spectra of AMY, ADH, GPDH, GDH, SDH, and EST are described and given in the paper. Enzyme systems, such as ADH, GPDH, and GDH, were found to be monomorphic. The EST enzyme showed the highest intracellular isoform diversity. Six obviously distinguished zones were found for this enzyme. Polymorphism is inherent for three of them.     

Isolation and characterization of poly(A)-containing polyoma "early" and "late" messenger RNAs.

During late lytic infection of mouse kidney cell cultures polyoma 16S and 19S (late 19S RNA) were isolated by oligo(dT)-cellulose chromatography. Approximately 60-80% of total cytoplasmic polyoma RNA contained tracts of poly(A) which were retained by oligo(dT)-cellulose. Early in lytic infection when viral DNA synthesis and the production of capsid protein are blocked by the addition of 5-fluorodeoxyuridine, approximately 100% of polyoma "early" 19S RNA was quantitatively retained by oligo(dT)-cellulose indicating the presence of poly(A) tracts on most 19S mRNA molecules. In addition, 2 classes polyoma RNA, synthesized after the onset of cellular RNA synthesis under conditions where DNA synthesis is inhibited with 5-fluorodeoxyuridine, were found to contain tracts of poly(A). These species sedimenting at 16S and 19S in aqueous sucrose density gradients were also quantitatively retained by oligo (dT)-cellulose.     

Identification of Long Non-Coding RNAs and Their Target Genes from Mycelium and Primordium in Model Mushroom Schizophyllum commune

Schizophyllum commune has emerged as the most promising model mushroom to study developmental stages (mycelium, primordium), which are two primary processes of fruit body development. Long non-coding RNA (lncRNA) has been proved to participate in fruit development and sex differentiation in fungi. However, potential lncRNAs have not been identified in S. commune from mycelium to primordium developmental stages. In this study, lncRNA-seq was performed in S. commune and 61.56 Gb clean data were generated from mycelium and primordium developmental stages. Furthermore, 191 lncRNAs had been obtained and a total of 49 lncRNAs were classified as differently expressed lncRNAs. Additionally, 26 up-regulated differently expressed lncRNAs and 23 down-regulated between mycelium and primordia libraries were detected. Further, Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that differentially expressed lncRNAs target genes from the MAPK pathway, phosphatidylinositol signal, ubiquitin-mediated proteolysis, autophagy, and cell cycle. This study provides a new resource for further research on the relationship between lncRNA and two developmental stages (mycelium, primordium) in S. commune.     

Comparison of developmentally controlled glucoamylase from normal and mutant strains of the basidiomycete Schizophyllum commune

During the formation of fruit bodies, the basidiomycete Schizophyllum commune produces glucoamylase activity. DEAE-cellulose chromatography resolves the enzymic activity into a major peak found early in development and a minor peak which appears when the fruit bodies are mature. A mutant homokaryon has been isolated which constitutively produces glucoamylase activity without any fruiting. When purified, the mutant enzyme and the major fruit body enzyme appear to be identical by several physical and kinetic parameters including molecular weight, temperature sensitivity, and K_m.Supported in part by NIH Research Grant AI 09779-03 from the National Institute of Allergy and Infectious Diseases.     

The modified nucleotide constituents of human prostatic cancer cell (MA-160) poly(A)-containing RNA

Cytoplasmic poly A(+) RNA from human prostatic cancer cells grown in the presence of 32P was isolated by affinity chromatography on columns of oligo(dT)-cellulose. The RNA was digested with RNAase T2 and the products of digestion were fractionated by two-dimensional electrophoresis. The resulting autoradiograms revealed the presence of many different cap groups as well as two internal modified nucleotide components. 19 different type 1 and type 2 'cap' groups were identified. The internal modified nucleotides were N6-methyl adenosine and a 2'-O-methyl nucleotide possessing an unusual modified base.     

Absence of short-period repetitive-sequence interspersion in the Basidiomycete Schizophyllum commune

Reassociation kinetics and electron microscopy were used to examine the organization of DNA sequences in the Basidiomycete Schizophyllum commune. Short-period interspersion of repetitive and unique sequences is absent from the DNA of this wood-rotting fungus. Instead, repetitive sequences are found predominantly in several, perhaps 16, clusters averaging 225 kilobases in length. The results of this study, the first concerning a fungus in the important fungal Class Basidiomycetidae, are discussed in relation to: sequence organization in three other species of true fungi, fungal evolution and the regulation of gene expression.