Shinji Ihara

Prefectural University of Hiroshima

The activation of ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) family proteases depends on removal of the prodomain. Although several studies suggest that ADAMTS activities play roles in development, homeostasis and disease, it remains unclear when and where the enzymes are activated in vivo. MIG-17, a Caenorhabditis elegan...

Large gaps in basement membrane occur at sites of cell invasion and tissue remodelling in development and cancer. Though never followed directly in vivo, basement membrane dissolution or reduced synthesis have been postulated to create these gaps. Using landmark photobleaching and optical highlighting of laminin and type IV collagen, we find that a...

Members of the ADAMTS (a disintegrin and metalloprotease with thrombospondin motifs) family of secreted proteins play important roles in animal development and pathogenesis. However, the lack of in vivo models has hampered elucidation of the mechanisms by which these enzymes are recruited to specific target tissues and the timing of their activatio...

A review of the Japanese

The migration of the gonadal distal tip cells (DTCs) in Caenorhabditis elegans provides an excellent model for studying the migration of epithelial tubes during organogenesis. Mutations in the mig-17/ADAMTS gene cause misdirected migration of DTCs during gonad formation, resulting in deformed gonad arms. An amino-acid substitution in RPL-20 corresp...

Figure 1 Localization patterns of mkate2::NID-1 and EMB-9::mcherry. (A and B) The localization of mkate2::NID-1 (right), and differential interference contrast (DIC) images (left) of nid-1(xyz8[mkate2::NID-1]) at L4 stage. (C

Figure 1. A. Sequence confirmation of the pign-1(xyz11) allele. Nucleotides and corresponding amino acid sequences in the wild-type pign-1 (upper) and xyz11 (bottom) are shown. The single guide RNA (sgRNA) target sequences are underlined in red, and the protospacer adjacent motif (PAM) is indicated by a red box. We used a single strand oligo, a hom...

Quality control of proteins in the endoplasmic reticulum (ER) is essential for ensuring the integrity of secretory proteins before their release into the extracellular space. Secretory proteins that fail to pass quality control form aggregates. Here we show the PIGN-1/PIGN is required for quality control in C. elegans and in mammalian cells. In C....

Overexpression of SPARC, a collagen-binding glycoprotein, is strongly associated with tumor invasion through extracellular matrix in many aggressive cancers. SPARC regulates numerous cellular processes including integrin-mediated cell adhesion, cell signaling pathways, and extracellular matrix assembly; however, the mechanism by which SPARC promote...

Quantification of SPARC overexpression. (A) Quantitative Real Time PCR of SPARC levels in wild type animals and animals expressing integrated transgenes overexpressing SPARC::GFP (syIs113 and syIs115). SPARC mRNA levels were standardized using act-4 (actin) mRNA as a control and normalized to wild type (N2) levels. Error bars denote standard deviat...

SPARC and type IV collagen colocalize in the ER. (A) SPARC::GFP (syIs115 line; left) and collagen::mCherry (center) colocalize in vesicles within the body wall muscle (overlay, right; average Pearson’s correlation coefficient = 0.70 ± 0.01; n = 8 animals). (B) Overexpressed SPARC::GFP (syIs115; left) is predominantly localized in the rough ER (as m...

SPARCR152L,Q159A::GFP, rab-3>SPARC::GFP, and SPARC::GFP localization at the BM. (A) hsp>SPARCR152L,Q159A::GFP (green, left; overlaid with DIC, right) is found in the BM (arrows). (B) Quantification of SPARC::GFP fluorescence at the BM in SPARC overexpression lines. As SPARC overexpression does not appear to affect the expression of endogenous unlab...

Time course of BM collagen levels after inducing SPARC overexpression. Worms expressing hsp>SPARC, hsp>SPARCR152L,Q159A::GFP, or no excess SPARC were subjected to a two hour heat shock at 32°C to drive SPARC overexpression, followed by four hours of recovery at 23°C. Collagen::mCherry fluorescence at the BM was measured at the indicated time points...

Characterization of two SPARC deletion alleles. Two non-overlapping deletions in the SPARC open reading frame (ost-1) were obtained from the C. elegans Gene Knockout Consortium. The location of the deletions tm6331 and tm966 is shown on top. Below, the progeny from a timed egg lay of ost-1(tm6331)/+ (left) or ost-1(tm966)/+ (right). The progeny of...

Overexpression of SPARC, a collagen-binding glycoprotein, is strongly associated with tumor invasion through extracellular matrix in many aggressive cancers. SPARC regulates numerous cellular processes including integrin-mediated cell adhesion, cell signaling pathways, and extracellular matrix assembly; however, the mechanism by which SPARC promote...

Microtubules (MTs) are cytoskeletal polymers that undergo dynamic instability, the stochastic transition between growth and shrinkage phases. MT dynamics are required for diverse cellular processes and, while intrinsic to tubulin, are highly regulated. However, little is known about how MT dynamics facilitate or are regulated by tissue biogenesis a...

The migration of Caenorhabditis elegans gonadal distal tip cells (DTCs) offers an excellent model to study the migration of epithelial tubes in organogenesis. mig-18 mutants cause meandering or wandering migration of DTCs during gonad formation, which is very similar to that observed in animals with mutations in mig-17, which encodes a secreted met...

α2-Heremans-Schmid glycoprotein (human fetuin) is one of numerous serum proteins produced in the liver. Recently, the biological functions of fetuin, such as calcification and insulin resistance, have been clarified. However, these effects appear to be indirect, occurring through binding to other molecules. When equal amounts of fetuin in sera were...

Integrin expression and activity have been strongly correlated with developmental and pathological processes involving cell invasion through basement membranes. The role of integrins in mediating these invasions, however, remains unclear. Utilizing the genetically and visually accessible model of anchor cell (AC) invasion in C. elegans, we have rec...

Glycosyltransferases are present in the Golgi apparatus in a membrane-bound form and are released from cells after cleavage by certain proteases. Beta1,6-N-acetylglucosaminyltransferase V (GnT-V), which is cleaved and secreted from the cells, is involved in the biosynthesis of beta1-6GlcNAc branching on N-glycans and has been implicated in tumor pr...

The core fucosylation (α1,6-fucosylation) of glycoproteins is widely distributed in mammalian tissues, and is altered under pathological conditions. To investigate physiological functions of the core fucose, we generated α1,6-fucosyltransferase (Fut8)-null mice and found that disruption of Fut8 induces severe growth retardation and death during pos...

Expression of N-acetylglucosaminyltransferase V (GnT-V) in colon cancer has been shown to be related to hematogenous metastasis and poor prognosis. To investigate the mechanism by which cancer cells expressing GnT-V metastasize to distant organs, we established GnT-V-overexpressing DLD-1 and WiDr cells (human colon cancer cell lines) by transfectin...

Multiple mechanisms are involved in the resistance of cancer cells to cisplatin, including the expression of multidrug resistance-associated protein (MRP) and enhanced DNA repair. Here, we report findings to show that oligosaccharide changes in alpha5beta1 integrin are associated with cisplatin resistance in a head and neck squamous cell carcinoma...

A glycomic approach to the identification of target molecules in glycosyltransferase gene targeting mice is a promising strategy to understand the biological significance of glycosyltransferase genes in vivo. In order to understand the biological effects of N-acetylglucosaminyltransferase III (GnT-III) on tumor formation in the liver, diethylnitros...

Ets-2, a transcriptional factor, has been linked to carcinoma progression but in-depth studies on its expression in human carcinoma have not been done. The present study investigated ets-2 expression in pancreatic adenocarcinoma. The ets-2 labeling index (LI) in normal pancreatic duct and pancreatic adenocarcinoma averaged 33.1+/-8.4 and 43.1+/-18....

Oligosaccharide moieties of glycoproteins are structurally altered during development, carcinogenesis, and malignant transformations. It is well known that β1–6 GlcNAc branching, a product of UDP-GlcNAc α-mannoside β1–6-N-acetylglucosaminyltransferase (GnT-V), is associated with malignant transformation as the results of such alterations. However,...

In the current concept of phototransduction, the concentration of cGMP in retinal rod outer segments is controlled by the balance of two enzyme activities: cGMP phosphodiesterase (PDE) and guanylyl cyclase (GC). However, no protein directly mediates these two enzyme systems. Here we show that RGS9, which is suggested to control PDE activity through...

In the current concept of phototransduction, the concentration of cGMP in retinal rod outer segments is controlled by the balance of two enzyme activities: cGMP phosphodiesterase (PDE) and guanylyl cyclase (GC). However, no protein directly mediates these two enzyme systems. Here we show that RGS9, which is suggested to control PDE activity through...