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The SUMO-conjugating enzyme Ubc9 is an essential enzyme in the SUMO (small ubiquitin-related modifier) protein modification system. Although sumoylation, covalent modification of cellular proteins by SUMO, is considered to regulate various cellular processes, and many substrates for sumoylation have been identified recently, the regulation of Ubc9 expression has not been examined in detail. We analyzed the expression of Ubc9 during rat brain development at the mRNA and protein levels. Northern and Western blot analyses revealed that expression of Ubc9 and SUMO-1 was developmentally regulated, while that of the ubiquitin-conjugating enzyme UbcH7 did not change so dramatically. In situ hybridization analysis revealed that the expression of Ubc9 was high in neuronal stem cells and moderate in differentiated neurons at embryonic stages. In the adult brain, moderate expression was observed in subsets of neurons, such as the dentate granular neurons and pyramidal neurons in the hippocampal formation and the large pyramidal neurons in the cerebral cortex. These results suggest that the Ubc9-SUMO system might participate in the proliferation and differentiation of neuronal cells in the developing brain and in neuronal plasticity in the adult brain.
We tested the reliability of fecal nitrogen (FN) to predict dietary nitrogen (DN) in two sika deerCervus nippon Heude, 1884 populations with greatly differing habitats. One was near the village of Nishiokoppe (area A) and the other was on Nakanoshima Island (area B) in Hokkaido, Japan. To estimate FN, we washed feces through a sieve, and diet was identified based on rumen-content analysis. The diet in area A was mostly composed of grass and legumes of agricultural origin, with browse being only a minor component. In contrast, browse such as deciduous foliage was the main component of the diet in area B. Dietary nitrogen was significantly regressed by FN within specific areas. On the other hand, the DN-FN-relationship had a similar slope but significantly different intercepts between areas. DN-FN-relationships differed between diets with and without an agricultural component, irrespective of browse. Thus, the difference in the DN-FN-relationship between areas was explained by differences in dietary composition. We therefore conclude that FN may be useful in predicting DN in diverse dietary situations regardless of the ratio of browse in the diet of free-ranging ungulates, but that dependence on agricultural pastures may hinder the utility of FN as an index of DN.
Previously, we demonstrated that when mouse erythroleukemia cells are exposed to a pressure of 80 MPa, the cell-cycle progression of S-phase cells is retarded. To examine the effects of high pressure on DNA replication, we used a Xenopus cell-free system. From cell-cycle progression of sperm nuclei, it was found that sperm nuclei are stable to a pressure of 80 MPa, whereas egg extracts are susceptible to high pressure. Similarly, biotin-16-dUTP was incorporated into 80 MPa-treated sperm nuclei in pressure-untreated extracts, but not into naive sperm nuclei in 80 MPa-treated extracts. These results indicate that DNA replication in Xenopus cell-free system is suppressed by the susceptibility of the extracts to a pressure of 80 MPa.
A comparative study on the utility of 2-(levulinyloxymethyl)-5-nitrobenzoyl (LMNBz) and 2-(levulinyloxymethyl)benzoyl (LMBz) protecting groups for the 5'-positions of nucleoside 3'-phosphoramidite derivatives in the oligonucleotide synthesis is presented in terms of the syntheses of TpTpT, TpTpTpT, and UpCpApGpUp-UpGpG. In addition we describe the synthesis, using the LMNBz protecting group, of the CpCpA terminus triplet of tRNAs bearing exocyclic amino groups with 15N-labeling, and the trimer Gp[A*]pG containing 2'-O-(β-D-ribofuranosyl)adenosine ([A*]), the latter of which is found at position 64 in the yeast initiator tRNAMet.
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