AtUBL5 regulates growth and development through pre-mRNA splicing in Arabidopsis thaliana

Ubiquitin-like proteins play important roles in the regulation of many biological processes. UBL5 (Ubiquitin-like protein 5)/Hub1 (Homologous to ubiquitin 1), a member of the ubiquitin family, acts as a ubiquitin-like modifier on a specific target, the spliceosomal protein Snu66, in yeast and human cells. The 22nd aspartic acid (Asp22) is involved in the attachment of Hub1 to the Hub1 interaction domain (HIND) of Snu66 in yeast to modulate spliceosomal activity. Hub1 differs from other modifiers which interact covalently with their targets. It modulates pre-mRNA splicing by binding to Snu66 non-covalently in both yeast and human cells. However, the molecular mechanisms of Hub1-mediated pre-mRNA splicing in plant systems remains unclear. To better understand the function of Hub1 in plants, we examined the role of this ubiquitin-like modifier in Arabidopsis thaliana, which has two Hub1 homologues. Arabidopsis UBL5/Hub1(UBL5) is highly conserved at the amino acid level, compared to eukaryotic homologues in both plants and animals. In this study, phenotypic analysis of A. thaliana with reduced UBL5 gene expression, generated by RNA interference of AtUBL5a and AtUBL5b were performed. Interestingly, knock down plants of AtUBL5 showed abnormalities in root elongation, plant development, and auxin response. AtUBL5b is highly expressed in the vascular tissue of the leaf, stem, and root tissue. Yeast two-hybrid analysis revealed that AtUBL5a and AtUBL5b interact with the putative splicing factor AtPRP38 through its C-terminal domain (AtPRP38C). Knock down of AtUBL5b resulted in a pattern of insufficient pre-mRNA splicing in several introns of AtCDC2, and in introns of IAA1, IAA4, and IAA5. Defects of pre-mRNA splicing in an AtPRP38 mutant resulted in an insufficient pre-mRNA splicing pattern in the intron of IAA1. Based on these results, we showed that AtUBL5b positively regulates plant root elongation and development through pre-mRNA splicing with AtPRP38C in A. thaliana

幼児期の過体重に影響する幼児の生活行動と家族の同居状況および生活行動

学位の種別: 論文博士審査委員会委員 : (主査)東京大学教授 水口 雅, 東京大学教授 上別府 圭子, 東京大学教授 渡辺 知保, 東京大学教授 橋本 英樹, 東京大学准教授 春名 めぐみUniversity of Tokyo(東京大学

Prevalence and characteristics of myocardial crypts in Japanese patients referred for cardiovascular magnetic resonance

18th Annual SCMR Scientific SessionsNice, France. 4-7 February 201

Quantitative Detection of µ Opioid Receptor: Western Blot Analyses Using µ Opioid Receptor Knockout Mice

Increasing evidence suggests that µ opioid receptor (MOP) expression is altered during the development of and withdrawal from substance dependence. Although anti-MOP antibodies have been hypothesized to be useful for estimating MOP expression levels, inconsistent MOP molecular weights (MWs) have been reported in studies using anti-MOP antibodies. In the present study, we generated a new anti-MOP antibody (N38) against the 1-38 amino acid sequence of the mouse MOP N-terminus and conducted Western blot analysis with wildtype and MOP knockout brain lysates to determine the MWs of intrinsic MOP. The N38 antibody detected migrating bands with relative MWs of 60-67 kDa in the plasma membrane fraction isolated from wildtype brain, but not from the MOP knockout brain. These migrating bands exhibited semi-linear density in the range of 3-30 µg membrane proteins/lane. The N38 antibody may be useful for quantitatively detecting MOP

Prevalence and characteristics of myocardial crypts in Japanese patients referred for cardiovascular magnetic resonance

18th Annual SCMR Scientific Sessions Nice, France. 4-7 February 201

Phenotypic Diversity and Altered Environmental Plasticity in <i>Arabidopsis thaliana</i> with Reduced Hsp90 Levels

The molecular chaperone HSP90 aids the maturation of a diverse but select set of metastable protein clients, many of which are key to a variety of signal transduction pathways. HSP90 function has been best investigated in animal and fungal systems, where inhibition of the chaperone has exceptionally diverse effects, ranging from reversing oncogenic transformation to preventing the acquisition of drug resistance. Inhibition of HSP90 in the model plant Arabidopsis thaliana uncovers novel morphologies dependent on normally cryptic genetic variation and increases stochastic variation inherent to developmental processes. The biochemical activity of HSP90 is strictly conserved between animals and plants. However, the substrates and pathways dependent on HSP90 in plants are poorly understood. Progress has been impeded by the necessity of reliance on light-sensitive HSP90 inhibitors due to redundancy in the A. thaliana HSP90 gene family. Here we present phenotypic and genome-wide expression analyses of A. thaliana with constitutively reduced HSP90 levels achieved by RNAi targeting. HSP90 reduction affects a variety of quantitative life-history traits, including flowering time and total seed set, increases morphological diversity, and decreases the developmental stability of repeated characters. Several morphologies are synergistically affected by HSP90 and growth temperature. Genome-wide expression analyses also suggest a central role for HSP90 in the genesis and maintenance of plastic responses. The expression results are substantiated by examination of the response of HSP90-reduced plants to attack by caterpillars of the generalist herbivore Trichoplusia ni. HSP90 reduction potentiates a more robust herbivore defense response. In sum, we propose that HSP90 exerts global effects on the environmental responsiveness of plants to many different stimuli. The comprehensive set of HSP90-reduced lines described here is a vital instrument to further examine the role of HSP90 as a central interface between organism, development, and environment.</p

Circulating inhibin and testosterone during sexual maturation and reproductive seasonality of captive male killer whales (Orcinus orca)

The present study aimed to investigate the reproductive biology of male killer whales. Changes in the concentrations of two circulating testicular hormones, inhibin and testosterone, were monitored during sexual maturation of two male Type 1 Eastern Northern Atlantic killer whales over a period of 20 years. The two killer whales grew rapidly at the pubertal stage and reached a plateau at the age of 23 and 20 years, respectively, after which growth slowed down. In the younger male, circulating inhibin was higher in the juvenile than in the pubertal and mature stages; whereas circulating testosterone exhibited the opposite trend. The pubertal period was estimated to last approximately 5 years, from 12 to 17 years of age. In the elder male, circulating testosterone was high from the onset of this study (12 years of age), when the animal also sired successfully for the first time. This finding shows that the male killer whale is possible to sire even if it is not socially matured, if there is opportunity for copulation. During the mature stage, both animals exhibited significantly higher circulating testosterone concentrations in spring compared to autumn and winter; whereas no seasonal change was observed for circulating inhibin. These results clearly demonstrate that the male killer whale is a seasonal breeder, even though it is fertile throughout the year. This is the first study to elucidate the inhibin concentration and secretory source in the male killer whale

Role of the Carboxy-Terminal Region of the GluRε2 Subunit in Synaptic Localization of the NMDA Receptor Channel

AbstractThe synaptic localization of the N-methyl-D-aspartate (NMDA) type glutamate receptor (GluR) channel is a prerequisite for synaptic plasticity in the brain. We generated mutant mice carrying the carboxy-terminal truncated GluRε2 subunit of the NMDA receptor channel. The mutant mice died neonatally and failed to form barrelette structures in the brainstem. The mutation greatly decreased the NMDA receptor–mediated component of hippocampal excitatory postsynaptic potentials and punctate immunofluorescent labelings of GluRε2 protein in the neuropil regions, while GluRε2 protein expression was comparable. Immunostaining of cultured cerebral neurons showed the reduced punctate staining of the truncated GluRε2 protein at synapses. These results suggest that the carboxy-terminal region of the GluRε2 subunit is important for efficient clustering and synaptic localization of the NMDA receptor channel

Patterns of Movement Performance Among Japanese Children and Effects of Parenting Practices: Latent class analysis

Objectives: The study aimed to examine the long-term effects of parenting practice during preschool years on children’s movement performance in primary school. Methods: This study involved a three-year longitudinal study including 225 children aged 3–6 years old. Parents reported baseline parenting practice and evaluated children’s movement performance three years later. Latent class analysis was used to explore latent classes of movement performance. A post hoc test was used to identify the characteristics of different patterns. Finally, adjusted multinomial logistic regression models were used to test the influence of parenting practice on identified patterns of movement performance. Results: Children in this study were grouped into three movement performance pattens, labelled as ‘least difficulties’ (58.2%, n = 131), ‘low back pain’ (30.2%, n = 68), and ‘most difficulties’ (11.6%, n = 26). After controlling for age, gender, having siblings or not, family structure, BMI SDS, sleep condition and dietary habits, we found that if parents played games with children frequently, the children would have a 0.287 times lower probability of being in the ‘low back pain’ class, 95%CI [0.105, 0.783], and if parents take children to meet peers of a similar age frequently, the children would have a 0.339 times lower probability of being in ‘most difficulties’ class, 95%CI [0.139, 0.825]. Conclusions: Primary healthcare providers should pay careful attention to children with movement difficulties. The study provides longitudinal evidence to support the applicability of positive parenting practice in early childhood to prevent children’s movement difficulties. Keywords: Movement performance; Parenting practice; Latent class analysis; Child; Longitudinal study; Japan

NF-kB阻害剤デヒドロキシメチルエポキシキノマイシン(DHMEQ)の鍵中間体および新規類縁体の合成

The synthesis of the key intermedicate of dehydroxymethylepoxyquinomicin (DHMEQ),a potent and specific NF-kB inhibitor,was achieved. The method involved the following crucial steps:i) direct construction of quinone functionality by degradative oxidation using hypervalent iodine (III) reagents such as (diacetoxyiodo) benzene (PIDA) and [bis(trifluoroacetoxy)iodo]beneze (PIFA);ii) regioselective epoxidation of quinone having a carbamoyl group; and iii) regio- and stereoselective reduction of an epoxyquinone moiety. In addition,a novel DHMEQ analog was synthesized by applying this approach