5 S RNA and 5.8 S RNA build up eukaryotic subribosomal domains active in tRNA binding.
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Biomedical subjects
Publications and source records attributed to M Saarma.
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Rat liver 5S rRNA and 5.8S rRNA were end-labelled with 32P at 5'-end or 3'-end of the polynucleotide chain and partially digested with single-strand specific S1 nuclease and double-strand specific endonuclease from the cobra Naja naja oxiana venom. The parallel use of these two structure-specific enzymes in combination with rapid sequencing technique allowed the exact localization of single-stranded and double-stranded regions in 5S RNA and 5.8 S RNA. The most accessible regions to S1 nuclease in 5S RNA are regions 33-42, 74-78, 102-103 and in 5.8 S RNA 16-20, 26-29, 34-36, 74-80 and a region around 125-130. The cobra venom endonuclease cleaves the following areas in 5S RNA: 7-8, 17-20, 28-30, 49-51, 56-57, 60-64, 69-70, 81-82, 95-97, 106-112. In 5.8S RNA the venom endonuclease cleavage sites are 4-7, 10-13, 21-22, 33-35, 43-45, 51-55, 72-74, 85-87, 98-99, 105-106, 114-115, 132-135. According to these results the tRNA binding sequences proposed by Nishikawa and Takemura [(1974) FEBS Lett. 40, 106-109], in 5S RNA are located in partly single-stranded region, but in 5.8S RNA in double-stranded region.
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A method for the isolation of structural genes, whose transcripts do not contain terminal poly(A) sequences, is presented. Poly(A) tails of predicted length were synthesized at the 3'-OH ends of RNA molecules employing Escherichia coli ATP : RNA adenyltransferase (EC 2.7.7.19). The gene isolation was performed in two steps: (a) enrichment of DNA fragments carrying the genes of interest, (b) subsequent cloning and amplification of these fragments. The enrichment of given gene sequences requires separate purifications of each of two complementary DNA chains, followed by their annealing. Rat liver 5S RNA genes were isolated by this method. DNA (+)chains were obtained by hybridizing EcoRI-digested and denatured rat liver DNA fragments with the 5S RNA-poly(A). The complex was purified on an oligo(dT)-cellulose column. DNA (-)chains were purified by hybridizing DNA fragments with the 5S cDNA which was coupled to oligo(dT)-cellulose. In the isolated and annealed double-stranded DNA preparation the enrichment of 5S DNA was 350--400-fold, when compared with the total genomic DNA. These purified DNA sequences were inserted into the EcoRI site of pBR325. Out of 1200 recombinant-plasmid colonies, analyzed in the colony-hybridization test, 4 hybridized with [125I]5S RNA.
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The distribution of oligonucleotides which are released from rat liver ribosomes by treatment with pancreatic ribonuclease has been studied. Rat liver monoribosomes lost from 15 to 17% of their nucleotides by treatment with pancreatic ribonuclease. This quantity was highly reproducible and did not depend significantly on the temperature (0-20 degrees C) and time (10-120 min) of incubation or on the concentration of enzyme (1:5000-1:50). Whereas the amounts of oligonucleotides liberated was 16%, it was shown by column chromatography that they consisted of 71% mononucleotides, 16% dinucleotides, 6% trinucleotides, 4% tetranucleotides and 2% pentanucleotides and that these oligonucleotides were enriched in uridine, containing approximately half of the uridine residues present in the high-molecular-weitht ribosomal RNA. The high molecular weight of the RNA from ribonuclease-treated ribosomes was preserved until it was heated; after heating, RNA fragments having sedimentation coefficients of 5 S and less were present. It is inferred that the olignucleotides are derived from pyrimidine-rich clusters located in single-stranded "hairpin" loops on the outside surface of the ribosome.
In 15 normal volunteers the action of succinic semialdehyde (SSA), and in 24 volunteers the effect of sodium succinate (SS), on the HNA have been investigated. As a result it was found that SSA brings about some enhancement of the excitatory process in cortical - mainly verbal -activities, accompanied by subjective improvement of memory and concentration. In the other hand, the administration of SS improves the stability of cortical excitatory process and its equilibrium with cortical inhibition; and enhances the connecting activity of the verbal system. These changes parallel the subjective elevation of mood and of general tone. In the basis of this study, one may suggest that succinic acid may be a psychoenergizer. The task of further investigations is to establish the range of therapeutic activity of these compounds.
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Glial cell line-derived neurotrophic factor (GDNF) was first discovered as a potent survival factor for midbrain dopaminergic neurons and was then shown to rescue these neurons in animal models of Parkinson's disease. GDNF is a more potent survival factor for dopaminergic neurons and the noradrenergic neurons of the locus coeruleus than other neurotrophic factors, and an almost 100 times more efficient survival factor for spinal motor neurons than the neurotrophins. The members of the GDNF family, GDNF, neurturin (NTN), persephin (PSP), and artemin (ART), have seven conserved cysteine residues with similar spacing, making them distant members of the transforming growth factor-beta (TGF-beta) superfamily. Like the members of the neurotrophin family, the GDNF-like growth factors belong structurally to the cysteine knot proteins. Like neurotrophins, GDNF family proteins are responsible for the development and maintenance of various sets of sensory and sympathetic neurons but, in addition, GDNF and NTN are also responsible for the development and survival of the enteric neurons, and NTN for parasympathetic neurons. All neurotrophins bind to the p75 low-affinity receptor, but their ligand specificity is determined by trk receptor tyrosine kinases. GDNF, NTN, PSP, and ART mediate their signals via a common receptor tyrosine kinase, Ret, but their ligand specificity is determined by a novel class of glycosylphosphatidylinositol (GPI)-anchored proteins called the GDNF family receptor alpha (GFR alpha). GDNF binds preferentially to GFR alpha1, NTN GFR alpha2, ART GRF alpha3, and PSP GFR alpha4 as a co-receptor to activate Ret. GFR alpha4 has until now been described only from chicken. Although the GDNF family members signal mainly via Ret receptor tyrosine kinase, there is recent evidence that they can also mediate their signals via GFR alpha receptors independently of Ret. The GDNF family of growth factors, unlike neurotrophins, has a well-defined function outside the nervous system. Recent transgenic and organ culture experiments have clearly demonstrated that GDNF is a mesenchyme-derived signaling molecule for the promotion of ureteric branching in kidney development. NTN, ART, and PSP are also expressed in the developing kidney, and NTN and PSP induce ureteric branching in vitro, but their true in vivo role in kidney morphogenesis is still unclear.