A novel diacylglycerol acyltransferase (DGAT2) is decreased in human psoriatic skin and increased in diabetic mice

Psoriasis is a skin disease with epidermal keratinocyte hyperproliferation and altered differentiation. To identify novel psoriasis-related genes, we investigated differentially expressed genes between normal and psoriatic skin. We identified a novel acyl CoA:diacylglycerol acyltransferase 2 (DGAT2) gene, which was decreased in human psoriatic skin. DGAT2 mRNA was expressed in sebaceous glands of normal human skin. DGAT2 protein was detected on endoplasmic reticulum. DGAT2 catalyzes the final step in the production of triglycerides and the accumulation of triglycerides in the tissues is considered to be related to insulin resistance. Therefore, we also investigated the expression of the DGAT2 gene in diabetic mice. DGAT2 mRNA was increased in the adipose, small intestine, and skeletal muscle in diabetic mice.     

DNA conjugates as novel functional oligonucleotides

Oligodeoxynucleotides with RNA cleavage activity 1) were conjugated with amines and peptides by solid phase fragment condensation (SPFC). It was found that 29 mer DNA enzyme conjugated with spermine at its 5'-end showed higher affinity to the target RNA sequence and 40 times higher activity of cleavage than native DNA enzyme. It is also to be noted that conjugate DNA enzymes showed increased resistance against nuclease digestion.     

In vitro establishment of tumorigenic human B-lymphoblastoid cell lines transformed by Epstein-Barr virus

We studied tumorigenic and phenotypic characteristics of pre- and postimmortal human B-lymphoblastoid cell lines (LCLs) transformed by Epstein-Barr virus (EBV): preimmortal LCLs showed low telomerase activity and a normal diploid karyotype while postimmortal LCLs showed much higher telomerase activity and maintained a clonal aneuploidic state. Among five postimmortal LCLs tested, LCLs N0005 and N6803 formed colonies in agar medium and showed marked aneuploidy, and N6803 was transplantable into nude mice indicating that it had a complete malignant phenotype, but all preimmortal LCLs and the remaining three postimmortal LCLs lacked these characteristics. The products of tumor suppresser genes, p16(INK4A) and pRb, were downregulated in these two LCLs, and the p53 gene was mutated in N0005 LCL. We believe these results showed for the first time that some postimmortal EBV-transformed LCLs can become tumorigenic, contrary to previous reports, and that these LCLs provide an in vitro model of tumorigenesis induced by EBV.     

Involvement of Fes/Fps tyrosine kinase in semaphorin3A signaling

Collapsin response mediator proteins (CRMPs)/TOAD64/Ulips/DRPs and CRAM have emerged as strong candidates for a role in semaphorin signaling. In this study we identified Fes/Fps (Fes) tyrosine kinase in the CRMP-CRAM complex and investigated whether Fes was involved in semaphorin3A (Sema3A) signaling. In COS-7 cells, the interaction between Fes and plexinA1 (PlexA1) and the tyrosine phosphorylation of PlexA1 by Fes were observed; however, these events were significantly attenuated by co-expression of neuropilin-1 (NP-1). Even with NP-1 co-expression, Sema3A was able to enhance the association of Fes with PlexA1 and Fes-mediated tyrosine phosphorylation of PlexA1, CRAM and CRMP2. Co-expression of Fes with PlexA1 exhibited COS-7 cell contraction activity, indicating that Fes can convert inactive PlexA1 to its active form, whereas combination of Fes/NP-1/PlexA1 or Fes kinase-negative mutants/PlexA1 did not alter cell morphology. Finally, Sema3A-induced growth cone collapse of dorsal root ganglion neurons was suppressed by expression of Fes kinase-negative mutants. Taken together, our findings suggest that Fes links Sema3A signals to CRMP-CRAM, and that NP-1 negatively regulates PlexA1 activation by Fes in resting condition.     

Symbiotic Bradyrhizobium japonicum Reduces N2O Surrounding the Soybean Root System via Nitrous Oxide Reductase

N₂O reductase activity in soybean nodules formed with Bradyrhizobium japonicum was evaluated from N₂O uptake and conversion of ¹⁵N-N₂O into ¹⁵N-N₂. Free-living cells of USDA110 showed N₂O reductase activity, whereas a nosZ mutant did not. Complementation of the nosZ mutant with two cosmids containing the nosRZDFYLX genes of B. japonicum USDA110 restored the N₂O reductase activity. When detached soybean nodules formed with USDA110 were fed with ¹⁵N-N₂O, they rapidly emitted ¹⁵N-N₂ outside the nodules at a ratio of 98.5% of ¹⁵N-N₂O uptake, but nodules inoculated with the nosZ mutant did not. Surprisingly, N₂O uptake by soybean roots nodulated with USDA110 was observed even in ambient air containing a low concentration of N₂O (0.34 ppm). These results indicate that the conversion of N₂O to N₂ depends exclusively on the respiratory N₂O reductase and that soybean roots nodulated with B. japonicum carrying the nos genes are able to remove very low concentrations of N₂O.     

Structure and Function of the Golgi Complex in Rice Cells (II. Purification and Characterization of Golgi Membrane-Bound Nucleoside Diphosphatase)

The gene coding for phytoene desaturase of the bacterium Erwinia uredovora (crtI) was inserted into the chromosome of the cyanobacterium Synechococcus PCC7942 strain R2-PIM8. For expression of crtI in the heterologous host, two constructs with different promoters were introduced into Synechococcus. In the first, crtI was fused to the 5[prime] region of the psbA gene of the xanthophycean microalga Bumilleriopsis filiformis. The second construct carried crtI inserted downstream of the neomycin phosphotransferase II gene (nptII) from the transposon Tn5. Expression of crtI under the control of the respective promoter was shown by immunodetection of the gene product. The functionality of the heterologously expressed phytoene desaturase CRTI in the transformants was demonstrated by enzymic assays. The transformants acquired very strong resistance toward the bleaching herbicide norflurazon.     

Structure and Function of the Golgi Complex in Rice Cells: Characterization of Golgi Membrane Glycoproteins

The structure and synthesis of the saccharide chains of Golgimembrane glycoproteins in suspension-cultured rice (Oryza sativaL.) cells were studied. Peanut lectin (PNA) and Ulex europaeuslectin-I (UEA-I) have high affinity for typical O-linked saccharidechains and both recognized the saccharide chains of rice Golgimembrane glycoproteins. These glycoproteins were also sensitiveto alkali and to O-glycanase. These results indicate that theGolgi membrane glycoproteins have O-linked saccharide chains.Brefeldin A, a specific inhibitor of Golgi-mediated secretion,induced morphological changes in Golgi complexes and preventedthe synthesis of the saccharide chains of the membrane glycoproteinsthat could be recognized by PNA and UEA-I. These glycoproteinswere typically localized in all compartments of the Golgi complex.Monensin can arrest the transport of secretory proteins frommedial to trans Golgi compartments but did not affect the formationand localization of the Golgi membrane glycoproteins. Tunicamycin,an inhibitor of the synthesis of N-linked saccharide chains,did not inhibit the synthesis of the saccharide chains of theseGolgi membrane glycoproteins. These results strongly suggestthat the synthesis of O-linked saccharide chains of Golgi membraneglycoproteins is initiated in the cis Golgi compartment. (Received September 24, 1992; Accepted June 4, 1993)     

A cDNA Encoding the 57 kDa Subunit of a Cytokinin-Binding Protein Complex from Tobacco: the Subunit Has High Homology to S-Adenosyl-L-Homocysteine Hydrolase

130 kDa cytokinin-binding protein complex (CBP130) is a majorcytokinin binding entity in tobacco leaves (Nicotiana sylvestris)and contains two protein species of 57 and 36 kDa [CBP57 andCBP36, Mitsui and Sugiura (1993) Plant Cell Physiol. 34: 543].We have determined partial amino acid sequences of CBP57 andisolated cDNAs encoding the protein from a tobacco cDNA libraryusing an oligonucleotide probe. Sequence analysis revealed significanthomology between CBP57 and S-adenosyl-L-homocysteine hydrolasefrom other organisms, which catalyzes the reversible hydrolysisof S-adenosyl-L-homocysteine, a methyltransferase inhibitor.CBP57 contains an additional sequence of 41 amino acids whichis not present in animal and slime mold Sadenosyl-L-homocysteinehydrolases. This additional sequence is also found in the parsleyand Rhodobacter enzymes, suggesting that it is unique to photosyntheticorganisms. CBP57 is encoded by more than one nuclear genes intobacco. Northern and western blot analyses revealed that thelevel of expression of the genes is high in roots and low inleaves. They are also expressed in cultured tobacco cells. Wediscuss the possibility that at least some of the physiologicaleffects of cytokinin are mediated through the control of methylation/demethylationby regulating the intracellular concentration of S-adenosyl-L-homocysteinevia the hydrolase. (Received June 24, 1993; Accepted August 14, 1993)