Appropriate expression of imprinted genes on mouse chromosome 12 extends development of bi-maternal embryos to term

Recently, we reported that the restored regulation of imprinted gene expression from two regions -H19 differentially methylated region (H19-DMR) and intergenic germline-derived DMR (IG-DMR) - is sufficient for accomplishing full-term development in mice. In the present study, we determined the developmental ability of the bi-maternal embryos (BMEs) containing the non-growing oocyte genome with the IG-DMR deletion (ng(Deltach12)) and fully-grown (fg) oocyte genome. Foetuses derived from ng(Deltach12)/fg BMEs were alive at E19.5 but could not survive further. Comparison with BMEs derived from Igf2+/- ng/fg genomes suggests that bi-allelic H19 expression might be involved in foetal development.     

Distribution of Japanese temperate bass, <Emphasis Type="Italic">Lateolabrax japonicus</Emphasis>, eggs and pelagic larvae in Ariake Bay

We collected eggs and larvae of the Japanese temperate bass, Lateolabrax japonicus, and present horizontal and temporal changes of distribution relative to development and growth during the species pelagic life history in Ariake Bay. Sampling was conducted from the inner to central region (11 sampling stations) of Ariake Bay using a plankton net (80 cm diameter, 0.5-mm mesh) from November 2000 to February 2001. Both eggs and larvae were collected most abundantly in mid-December. The CPUE of eggs in the surface layer was higher than the middle layer, which is in contrast to that at the larval stage. Most eggs were collected around the central and western regions of the bay. The distribution of eggs shifted vertically to the middle layer with development. Yolk-sac larvae were collected in the central region of the bay, and preflexion and flexion larvae were more abundantly collected in the inner region of the bay. The body length of larvae around the inner bay was larger than in the central region. The pelagic life history can be summarized as follows: eggs are distributed around the central region of the bay and eggs and larvae expand their distribution to the inner and shallower waters with growth. We conclude that the shift of vertical distribution in pelagic stages and the hydrographic features of the middle layer form one of the mechanisms enabling the inshore migration of L. japonicus.     

Normal human chromosome 5, on which a familial adenomatous polyposis gene is located, has tumor suppressive activity.

The suppressive activity of normal human chromosome 5 was detected by means of the chromosomal transfer technique using DT cells as recipients. A hybrid clone, which exhibited reduced tumorigenicity, contained chromosomal regions such as 5pter-p15, q21 and q33-qter. Since a familial adenomatous polyposis gene has been reported to be located at 5q21-q22, the suppressive activity of chromosome 5 might be due to this gene.     

Isolation and characterization of an asparagine-linked keratan sulfate from the skin of a marine teleost, Scomber japobicus

Keratan sulfate was isolated from the skin of Pacific mackerel (Scomber japonicus) after exhaustive digestion with pronase followed by ethanol precipitation and fractionation on a cellulose column with 0.3% recovery of dried material. The keratan sulfate preparation was separated into four major fractions by Dowex-1 column chromatrography. The chemical and infrared spectrum analyses of the four fractions showed a high degree of heterogeneity in sulfation. Since the carbohydrate-peptide linkage in the teleost skin keratan sulfate was found to be stable in alkali, and asparagine was the predominant amino acid, the asparagine residue in the peptide backbone was most likely to be involved in the N-glycosyl linkage with the carbohydrate moiety. Besides the type of carbohydrate-peptide linkage, the teleost skin keratan sulfate is very similar to corneal keratan sulfate, (keretan sulfate I) in two respects: (1) The teleost skin and bovine corneal keratan sulfates were hydrolyzed much faster by endo-β-galactosidase that the whale nasal cartilage keratan sulfate (keratan sulfate II). (2) Although the teleost skin keratan sulfate showed considerable polydispersity, the molecular weight was in the same range as the corneal keratan sulfate, and it was relatively higher than that of the cartilage keratan sulfate.     

Phenylalanine 171 is a molecular brake for translesion synthesis across benzo[a]pyrene-guanine adducts by human DNA polymerase kappa

Human cells possess multiple specialized DNA polymerases (Pols) that bypass a variety of DNA lesions which otherwise would block chromosome replication. Human polymerase kappa (Pol κ) bypasses benzo[a]pyrene diolepoxide-N(2)-deoxyguanine (BPDE-N(2)-dG) DNA adducts in an almost error-free manner. To better understand the relationship between the structural features in the active site and lesion bypass by Pol κ, we mutated codons corresponding to amino acids appearing close to the adducts in the active site, and compared bypass efficiencies. Remarkably, the substitution of alanine for phenylalanine 171 (F171), an amino acid conserved between Pol κ and its bacterial counterpart Escherichia coli DinB, enhanced the efficiencies of dCMP incorporation opposite (-)- and (+)-trans-anti-BPDE-N(2)-dG 18-fold. This substitution affected neither the fidelity of TLS nor the efficiency of dCMP incorporation opposite normal guanine. This amino acid change also enhanced the binding affinity of Pol κ to template/primer DNA containing (-)-trans-anti-BPDE-N(2)-dG. These results suggest that F171 functions as a molecular brake for TLS across BPDE-N(2)-dG by Pol κ and that the F171A derivative of Pol κ bypasses these DNA lesions more actively than does the wild-type enzyme.