A5043318623- Plant Virus Research Studies
- Plant and Fungal Interactions Research
- RNA and protein synthesis mechanisms
- Enzyme Production and Characterization
- Insect-Plant Interactions and Control
- Enzyme Structure and Function
- RNA modifications and cancer
- Biofuel production and bioconversion
- Polyamine Metabolism and Applications
- Peptidase Inhibition and Analysis
- Bacteriophages and microbial interactions
- Fungal Plant Pathogen Control
- CRISPR and Genetic Engineering
- Plant Reproductive Biology
- Glycosylation and Glycoproteins Research
- Insect and Pesticide Research
- Microbial Metabolites in Food Biotechnology
- Polysaccharides and Plant Cell Walls
- Mosquito-borne diseases and control
- Plant Parasitism and Resistance
- Transgenic Plants and Applications
- Plant Molecular Biology Research
- Genomics and Chromatin Dynamics
- Insect Resistance and Genetics
- Advanced Proteomics Techniques and Applications
Hokkaido University
2004-2017
Institute of Agrobiological Sciences
2006-2014
Japan Science and Technology Agency
2004
α-Glucosidases, which catalyze the hydrolysis of α-glucosidic linkage at nonreducing end substrate, are important for metabolism α-glucosides. Halomonas sp. H11 α-glucosidase (HaG), belonging to glycoside hydrolase family 13 (GH13), only has high hydrolytic activity towards α-(1 → 4)-linked disaccharide maltose among naturally occurring substrates. Although several three-dimensional structures GH13 members have been solved, specificity and 4) recognition mechanism unclear owing a lack...
The replication of eukaryotic positive-strand RNA virus genomes occurs through a complex process involving multiple viral and host proteins intracellular membranes. Here we report cell-free system that reproduces this in vitro . This uses membrane-containing extract uninfected plant protoplasts from which the vacuoles had been removed by Percoll gradient centrifugation. We demonstrate supported translation, negative-strand synthesis, genomic replication, subgenomic transcription tomato...
Abstract RNA viruses use various strategies to condense their genetic information into small genomes. Potyviruses not only the polyprotein strategy, but also embed an open reading frame, pipo , in P3 cistron –1 frame. PIPO is expressed as a fusion protein with N-terminal half of (P3N-PIPO) via transcriptional slippage viral RNA-dependent polymerase (RdRp). We herein show that clover yellow vein virus (ClYVV) produces previously unidentified factor, P3N-ALT, +1 frame at conserved G 1–2 A 6–7...
Proteins Rpf2 and Rrs1 are required for 60S ribosomal subunit maturation. These proteins necessary the recruitment of three components (5S RNA [rRNA], RpL5 RpL11) to 90S ribosome precursor subsequent 27SB pre-rRNA processing. Here we present crystal structure Aspergillus nidulans (An) Rpf2-Rrs1 core complex. The complex contains tightly interlocked N-terminal domains Rrs1. domain includes a Brix characterized by similar N- C-terminal architecture. long α-helix joins half as if it were part...
Tomato spotted wilt virus (TSWV), belonging to the genus Tospovirus of family Bunyaviridae, causes significant economic damage several vegetables and ornamental plants worldwide. Similar those all other negative-strand RNA viruses, nucleocapsid (N) protein plays very important roles in its viral life cycle. N proteins protect genomic RNAs by encapsidation form a ribonucleoprotein complex (vRNP) with some RNA-dependent polymerases. Here we show crystal structure from TSWV. Protomers TSWV...
The replication of eukaryotic positive-strand RNA virus genomes occurs in the membrane-bound complexes. Previously, we found that extract evacuolated tobacco BY-2 protoplasts (BYL) is capable supporting translation and subsequent genomic RNAs plant viruses, including Tomato mosaic (ToMV). Here, to dissect process precedes formation ToMV complexes, prepared membrane-depleted BYL (mdBYL), which membranes were removed by centrifugation. In mdBYL, was translated produce 130-kDa 180-kDa proteins,...
The analysis of post-transcriptional regulatory mechanisms in plants has benefited greatly from the use cell-free extract systems. Arabidopsis as a model system provides extensive genetic resources; however, to date suitable translation not been available. In this study, we devised an (ACE) be used for vitro studies. Protoplasts were prepared callus cultures derived seedlings, and extracts after evacuolation protoplasts by Percoll gradient centrifugation. new ACE exhibits activity comparable...
Significance Template-dependent RNA and DNA polymerization is a vital reaction in the cell believed to occur exclusively forward direction (5′-3′), which poses significant challenges in, for example, lagging strand synthesis. Although cells are mostly limited unidirectional polymerization, we find that reverse structurally chemically possible utilizing same structural core, conserved palm domain of canonical polymerases. The structure unique nucleotide polymerase-tRNA complex revealed...
In Ruminococcus albus , 4‐ O ‐β‐ d ‐mannosyl‐ ‐glucose phosphorylase ( Ra MP 1) and β‐(1,4)‐mannooligosaccharide 2) belong to two subfamilies of glycoside hydrolase family 130. The enzymes phosphorolyze β‐mannosidic linkages at the nonreducing ends their substrates, have substantially diverse substrate specificity. differences in mechanism binding not yet been fully clarified. present study, we report crystal structures 1 with/without 2 β‐(1→4)‐mannobiose. differ +1 subsite substrate‐binding...
Rpf2 and Rrs1 are essential proteins for ribosome biogenesis. These form a complex (the Rpf2-subcomplex) with 5S rRNA two ribosomal (L5 L11). This is recruited to the precursor 90S pre-ribosome). recruitment necessary maturation of 25S rRNA. Genetic depletion results in accumulation precursor. In this study, from Aspergillus nidulans were co-overexpressed Escherichia coli, purified crystallized. Subsequent analysis revealed that these crystals contained central core region consisting both...
The α-glucosidase HaG from the halophilic bacterium Halomonas sp. strain H11 catalyzes hydrolysis of glucosidic linkage at nonreducing end α-glucosides, such as maltose and sucrose, to release α-glucose. Based on its amino-acid sequence, this enzyme is classified a member glycoside hydrolase family 13. has three unique characteristics: (i) very narrow substrate specificity, almost exclusively hydrolyzing disaccharides; (ii) activation by monovalent cations, K(+), Rb(+), Cs(+) NH4(+); (iii)...
Tomato spotted wilt virus (TSWV), which causes severe damage to various agricultural crops such as tomato, pepper, lettuce and peanut, is a negative-stranded RNA belonging the Tospovirus genus of Bunyaviridae family. Viral genomic molecules are packaged in form ribonucleoprotein complexes, each contains one viral molecule that coated with many nucleocapsid (N) proteins. Here, expression crystallization TSWV N protein reported. Native selenomethionine-substituted crystals belonged same space...
Recent progress in the techniques of bio-macromolecular crystallography makes crystal structure analysis more powerful and useful for life science. The huge super-molecular (eukaryotic Ribosome, Vault etc.) membrane proteins related to diseases were successful. Moreover, structure/fragment drug design using method is also becoming reliable. However, crystallization still remains as a major bottleneck determining structures, although many methods have been developed such kits, robot,...