Activin in the brain modulates anxiety-related behavior and adult neurogenesis

Activin, a member of the transforming growth factor-beta superfamily, is an endocrine hormone that regulates differentiation and proliferation of a wide variety of cells. In the brain, activin protects neurons from ischemic damage. In this study, we demonstrate that activin modulates anxiety-related behavior by analyzing ACM4 and FSM transgenic mice in which activin and follistatin (which antagonizes the activin signal), respectively, were overexpressed in a forebrain-specific manner under the control of the alphaCaMKII promoter. Behavioral analyses revealed that FSM mice exhibited enhanced anxiety compared to wild-type littermates, while ACM4 mice showed reduced anxiety. Importantly, survival of newly formed neurons in the subgranular zone of adult hippocampus was significantly decreased in FSM mice, which was partially rescued in ACM4/FSM double transgenic mice. Our findings demonstrate that the level of activin in the adult brain bi-directionally influences anxiety-related behavior. These results further suggest that decreases in postnatal neurogenesis caused by activin inhibition affect an anxiety-related behavior in adulthood. Activin and its signaling pathway may represent novel therapeutic targets for anxiety disorder as well as ischemic brain injury.     

Promoter exchange between ompF and ompC, genes for osmoregulated major outer membrane proteins of Escherichia coli K-12.

Expression of the ompF and ompC genes coding for major outer membrane proteins OmpF and OmpC is regulated in opposite directions by medium osmolarity. Chimera genes were constructed by a reciprocal exchange of the promoter-signal sequence region between the two genes. The chimera gene construction was designed so that the proteins synthesized by these genes were essentially the same as the OmpC and OmpF proteins. Studies with the chimera genes demonstrated that the osmoregulation of the OmpF-OmpC synthesis was promoter dependent. They also showed that cells grew normally even when the osmoregulation took place in opposite directions. The effects of the ompR2 and envZ mutations, which suppress ompC and ompF expression, respectively, also became reversed. The reduced expression was still subject to the promoter-controlled osmoregulation. Based on these observations, the mechanism of regulation of the ompF-ompC gene expression and its physiological importance are discussed.     

Developmental morphology of one-leaf plant Monophyllaea singularis (Gesneriaceae)

 Developmental morphology is described of the one-leaf plant Monophyllaea singularis which possesses a huge macrocotyledon, a long petiolode below it, and many small inflorescences scattered along the petiolode and midrib. Cell proliferation and basipetal differentiation occur in both cotyledons after water imbibition and germination. The basal meristem forms from a group of small, least differentiated cells at the base of a future macrocotyledon and continues blade production even at the reproductive stage. The petiolode meristem, which forms as an intercalary meristem near the base of the macrocotyledon, contributes to the elongation of the petiolode and the midrib. Although the 'groove meristem', like the groove meristem of Streptocarpus, forms between the cotyledons at the site of a shoot apical meristem, it is not involved in inflorescence production. In M. singularis, instead of the 'groove meristem', the inflorescences are initiated adventitiously from groups of cells in the dermal and subdermal layers of the petiolode and probably also of the midrib. Received October 10, 2000 Accepted August 2, 2001     

Crystallization and preliminary crystallographic analysis of a 2,3-dihydroxybiphenyl dioxygenase from Pseudomonas sp. strain KKS102 having polychlorinated biphenyl (PCB)-degrading activity

Crystals have been obtained for a 2,3-dihydroxybiphenyl dioxygenase (conventionally called BphC) from a polychlorinated biphenyl (PCB)-degrader, Pseudomonas sp. strain KKS1O2. The crystals were grown using both ammonium sulfate and MPD as the precipitating agents. The crystals belonged to a tetragonal space group (I422) and diffracted to 2.5 Å. © 1995 Wiley-Liss, Inc.     

Differential expression of two hemA mRNAs encoding glutamyl-tRNA reductase proteins in greening cucumber seedlings.

The first committed step of porphyrin synthesis in higher plants is the reduction of glutamyl-tRNA to glutamate 1-semialdehyde. This reaction is catalyzed by glutamyl-tRNA reductase, which is encoded by hemA genes. Two hemA cDNA clones (hemA1 and hemA2) were obtained from cucumber (Cucumis sativus) cotyledons by the PCR and cDNA library screening. They showed significant homology with published hemA sequences. Southern blot analysis of cucumber genomic DNA revealed that these genes are located at different loci and that there is another gene similar to the hemA genes. Accumulation of hemA1 mRNA was detected primarily in cotyledons and hypocotyls of greening cucumber seedlings, whereas that of hemA2 mRNA was detected in all tissues examined. Illumination of cucumber seedlings increased markedly the accumulation of hemA1 mRNA, but it did not induce remarkable changes in that of hemA2 mRNA. These findings suggest that hemA1 mRNA was accumulated in response to the demand of Chl synthesis in photosynthesizing tissues, whereas hemA2 mRNA was expressed in response to the demand of the synthesis of porphyrins other than chlorophylls.     

Genetic analysis of functional connectivity between substrate recognition domains ofEscherichia coli glutaminyl-tRNA synthetase

It has previously been shown that the single mutation E222K in glutaminyl-tRNA synthetase (GlnRS) confers a temperature-sensitive phenotype onEscherichia coli. Here we report the isolation of a pseudorevertant of this mutation, E222K/C171G, which was subsequently employed to investigate the role of these residues in substrate discrimination. The three-dimensional structure of the tRNA^Gln: GlnRS:ATP ternary complex revealed that both E222 and C171 are close to regions of the protein involved in interactions with both the acceptor stem and the 3′ end of tRNA^Gln. The potential involvement of E222 and C171 in these interactions was confirmed by the observation that GlnRS-E222K was able to mischargesupF tRNA^Tyr considerably more efficiently than the wild-type enzyme, whereas GlnRS-E222K/C171G could not. These differences in substrate specificity also extended to anticodon recognition, with the double mutant able to distinguishsupE tRNA _CUA ^Gln from tRNA ₂ ^Gln considerably more efficiently than GlnRS E222K. Furthermore, GlnRS-E222K was found to have a 15-fold higher K_m for glutamine than the wild-type enzyme, whereas the double mutant only showed a 7-fold increase. These results indicate that the C171G mutation improves both substrate discrimination and recognition at three domains in GlnRS-E222K, confirming recent proposals that there are extensive interactions between the active site and regions of the enzyme involved in tRNA binding.     

Mutation causing reverse osmoregulation of synthesis of OmpF, a major outer membrane protein of Escherichia coli.

Supplementation of growth media with high concentrations of substances like sucrose results in the induction of OmpC synthesis and the suppression of OmpF synthesis. We isolated a novel mutant in which OmpF synthesis is in the opposite direction from normal osmoregulation. By transductional mapping, the mutation was localized at 75 min between malA and aroB on the Escherichia coli chromosome map where the ompR-envZ region is. The mutation was suppressed by a plasmid carrying the ompR gene but not by a plasmid carrying the envZ gene alone. The mutation also resulted in the almost complete suppression of OmpC synthesis. However, the remaining OmpC synthesis was osmoregulated normally. Based on these observations, the mechanism of osmoregulation of OmpF-OmpC synthesis is discussed.     

Kinetic study on the successive four-step reduction of Cyt c3

A detailed kinetic study on the successive four-step reduction of cyt c3, which has four heme units in a single protein, III4 leads to III3II leads to III2II2 leads to III II3 leads to II4, was carried out by stopped-flow electronic spectroscopy (SF-UV) and stopped-flow circular dichroism spectroscopy (SF-CD). Based on the absorbance change vs. time and the ellipticity change vs. time at the characteristic CD, together with the electronic absorption of the enzyme, rate constants for the successive four electron transfer steps, k1-k4, were successfully estimated by computer simulation. The rate constants of the four steps (k1 = 19.8 s-1, k2 = 11.9 s-1, k3 = 8.9 s-1, and k4 = 1.6 s-1; 8.0 10(-4) M Na2S2O4) are quite different from the statistical values (4: 3: 2: 1), thus excluding the possibility of random reduction of hemes of equal reactivities. Instead, each heme has its own reactivity, probably dependent on its local environment. The value of k3 is somewhat higher than the statistical value, indicating the existence of an autoacceleration effect, although small. This autoacceleration is most probably due to a unique heme-heme and/or heme-environment interaction since unusual CD and electronic absorptions were observed at 350-400 nm at about the time corresponding.     

Hydroxyurea inhibits degradation of internalized epidermal growth factor in HeLa cells

Because epidermal growth factor stimulates DNA synthesis in cultured cells, five inhibitors of DNA synthesis were tested in HeLa cells to see whether the inhibition of DNA synthesis has any effect on the metabolism of the growth factor. Among these, only hydroxyurea inhibited the degradation of ¹²⁵I-labeled epidermal growth factor strongly. The reversal of hydroxyurea-induced inhibition of DNA synthesis by deoxyribonucleosides did not result in a recovery from the inhibition of the degradation. From these findings, it might be concluded that the inhibitory effect of hydroxyurea on the degradation is distinct from that on DNA synthesis.