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Biomedical subjects

Yoichi Noda

Publications and source records attributed to Yoichi Noda.

At least 37 records · Page 2Linked to original sources
PubMed2005

ROCK-I regulates closure of the eyelids and ventral body wall by inducing assembly of actomyosin bundles.

Rho-associated kinase (ROCK) I mediates signaling from Rho to the actin cytoskeleton. To investigate the in vivo functions of ROCK-I, we generated ROCK-I-deficient mice. Loss of ROCK-I resulted in failure of eyelid closure and closure of the ventral body wall, which gave rise to the eyes open at birth and omphalocele phenotypes in neonates. Most ROCK-I(-/-) mice died soon after birth as a result of cannibalization of the omphalocele by the mother. Actin cables that encircle the eye in the epithelial cells of the eyelid were disorganized and accumulation of filamentous actin at the umbilical ring was impaired, with loss of phosphorylation of the myosin regulatory light chain (MLC) at both sites, in ROCK-I(-/-) embryos. Stress fiber formation and MLC phosphorylation induced by EGF were also attenuated in primary keratinocytes from ROCK-I(-/-) mice. These results suggest that ROCK-I regulates closure of the eyelids and ventral body wall through organization of actomyosin bundles.

Abdominal Wall↗
PubMed2005

SKG1, a suppressor gene of synthetic lethality of kex2Deltagas1Delta mutations, encodes a novel membrane protein that affects cell wall composition.

The fungal GAS1-related genes encode GPI-anchored beta-1,3-glucanosyltransferase, and their loss causes a defect in the assembly of the cell wall. The KEX2 gene encodes a processing protease in the late Golgi compartment and its loss also results in defects in the cell wall. Simultaneous mutations of these genes are lethal in Saccharomyces cerevisiae. To understand the basis of this synthetic lethality, we screened for multicopy suppressors and identified 13 SKG (suppressor of kex2 gas1 synthetic lethality) genes. SKG1 encodes a transmembrane protein that localizes on the inner surface of the plasma membrane at the bud and in the daughter cell. The multicopy SKG1 increases the sensitivity of cells to zymolyase, and the skg1Delta null mutation increases resistance to it. This zymolyase susceptibility corresponds to an increase of alkali-soluble beta-1,3-glucan and a decrease of chitin in the cell wall. Thus SKG1 encodes a novel protein that affects the cell wall polymer composition in the growing region of the cell.

Amino Acid Sequence↗
PubMed2005

Effectiveness of two-step (consecutive) embryo transfer in patients who have two embryos on day 2: comparison with cleavage-stage embryo transfer.

OBJECTIVE: To evaluate the effectiveness, for patients who have only two embryos on day 2, of a two-step (consecutive) embryo transfer (ET) procedure in which a cleaved embryo is transferred on day 2 and a single blastocyst is transferred on day 5. DESIGN: Observational comparative study. SETTING: Private IVF clinic. PATIENT(S): Ninety two-step ET cycles were performed in patients who had two embryos on day 2 (two-step group). Ninety day-2 ET cycles were performed in age- and infertility-matched patients who had two embryos on day 2 (control group). INTERVENTION(S): Cleaved-ET, extended culture of one embryo, and a second transfer of a blastocyst. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): The pregnancy and implantation rates in the two-step group (respectively 33.3% and 17.2%) were significantly higher than those in the control group (18.9% and 9.4%). Thirty-nine of the patients in the two-step group (43.3%) could not proceed to the second step of ET because no viable blastocyst could be obtained, but four of them conceived anyway. CONCLUSION(S): Taking advantage of both day-2 ET and blastocyst transfer, two-step ET may be an effective option for ET in patients who have an insufficient number of embryos.

Adult↗
PubMed2005

Immunoisolaton of the yeast Golgi subcompartments and characterization of a novel membrane protein, Svp26, discovered in the Sed5-containing compartments.

The Golgi apparatus consists of a set of vesicular compartments which are distinguished by their marker proteins. These compartments are physically separated in the Saccharomyces cerevisiae cell. To characterize them extensively, we immunoisolated vesicles carrying either of the SNAREs Sed5 or Tlg2, the markers of the early and late Golgi compartments, respectively, and analyzed the membrane proteins. The composition of proteins was mostly consistent with the position of each compartment in the traffic. We found six uncharacterized but evolutionarily conserved proteins and named them Svp26 (Sed5 compartment vesicle protein of 26 kDa), Tvp38, Tvp23, Tvp18, Tvp15 (Tlg2 compartment vesicle proteins of 38, 23, 18, and 15 kDa), and Gvp36 (Golgi vesicle protein of 36 kDa). The localization of Svp26 in the early Golgi compartment was confirmed by microscopic and biochemical means. Immunoprecipitation indicated that Svp26 binds to itself and a Golgi mannosyltransferase, Ktr3. In the absence of Svp26, a considerable portion of Ktr3 was mislocalized in the endoplasmic reticulum. Our data suggest that Svp26 has a novel role in retention of a subset of membrane proteins in the early Golgi compartments.

Amino Acid Sequence↗
PubMed2004

A novel endoplasmic reticulum membrane protein Rcr1 regulates chitin deposition in the cell wall of Saccharomyces cerevisiae.

Congo red binds to the cell wall and inhibits the growth of yeast. In a screening for multicopy suppressor genes of Congo red hypersensitivity of erd1Delta mutant, we found that a previously uncharacterized gene, YBR005w, makes most of the Saccharomyces cerevisiae strains resistant to Congo red. This gene was named RCR1 (resistance to Congo red 1). An rcr1Delta null mutant showed an increased sensitivity to Congo red. RCR1 encodes a novel ER membrane protein with a single transmembrane domain. Molecular dissection suggested that the transmembrane domain and a part of the C-terminal polypeptide are sufficient for the activity. We examined the effect of RCR1 in various null mutants of genes related to the cell wall. The resistance of mutants to Congo red correlates with a reduction of chitin content. Multicopy RCR1 caused a significant decrease in the chitin content while the amount of alkali-soluble glucan did not change. The binding of Calcofluor white to the cell wall significantly decreased in these cells. Our results show that RCR1 regulates the chitin deposition and add firm genetic and biochemical evidences that the primary target of Congo red is chitin in S. cerevisiae.

Amino Acid Sequence↗
PubMed2004

Localization of GDP-mannose transporter in the Golgi requires retrieval to the endoplasmic reticulum depending on its cytoplasmic tail and coatomer.

The Saccharomyces cerevisiae GDP-mannose transporter (GMT) encoded by the essential gene VRG4/VIG4 is a member of the nucleotide-sugar transporter family in the Golgi apparatus. We examined GMT in the secretory mutant cells to investigate the mechanism of its localization in the Golgi. At the nonpermissive temperature, most GMT was found in the endoplasmic reticulum of sec23ts cells, which have defective COPII, and in the vacuole of sec21ts cells, which have defective COPI. The C-terminal hydrophilic peptide of GMT that is exposed to the cytosol binds to Ret2p, a subunit of the COPI coat. Mutant peptide derivatives that have lost a cluster of lysine in the vicinity of the transmembrane domain had reduced binding activity to Ret2p and the GMT with this sequence was delivered to the vacuole. Our results indicate that GMT escapes from delivery to the vacuole by recycling to the endoplasmic reticulum and retrieval requires the lysine-rich C-terminal tail that can bind to the COPI coat.

Amino Acid Sequence↗
PubMed2003

In utero diagnosis of an aneurysm of the vein of Galen causing hydrocephalus and heart failure.

A prenatally diagnosed aneurysm of the vein of Galen was presented in the fetus of a patient referred to our hospital at 31 weeks of gestation. Ultrasonography demonstrated polyhydramnios, cardiomegaly, dilatation of the right atrium and superior vena cava, tricuspid valve regurgitation, hydrocephalus, and a large hypoechoic mass with blood flow in the suboccipital region. Skin edema was shown thereafter. A 3262-g male was delivered by cesarean at 35 weeks of gestation. Computed tomography imaging demonstrated a large mass in the suboccipital region, after which thrombocytopenia appeared and the neonate died at 18 days of age.

Adult↗
PubMed2003

Follistatin-related gene (FLRG) expression in human endometrium: sex steroid hormones regulate the expression of FLRG in cultured human endometrial stromal cells.

Follistatin-related gene (FLRG) encodes a novel secreted glycoprotein that is highly homologous to follistatin and binds activins and bone morphogenetic proteins, members of the TGF beta superfamily of growth/differentiation factors. FLRG protein inhibits activin-induced and bone morphogenetic protein-2-induced transcriptional responses in a dose-dependent manner, and its mRNA is abundantly expressed in human placenta, heart, aorta, testis, and adrenal gland. In this study we showed that FLRG mRNA was expressed in human endometrium across the menstrual cycle and in decidua of early pregnancy. In the proliferative phase of the menstrual cycle, FLRG protein was detected predominantly in the cytoplasm of endometrial epithelium. In the secretory phase and in early pregnancy, it was also detected in the nuclei of endometrial stromal cells. Using in vitro decidualization model, we demonstrated that 17 beta-estradiol plus progesterone, but not 17 beta-estradiol or progesterone alone, induced FLRG expression significantly. These results suggest that FLRG expression in endometrial stromal cells is regulated by the concerted action of ovarian steroid hormones via decidualization, and FLRG protein may participate in the regulation of stromal cell decidualization as a binding protein for members of TGF beta superfamily.

Adult↗
PubMed2003

Cooperation of Sly1/SM-family protein and sec18/NSF of Saccharomyces cerevisiae in disassembly of cis-SNARE membrane-protein complexes.

Assembly and disassembly of the SNARE membrane-protein complexes plays a key role in vesicular trafficking. The SM-family Slyl protein binds to the tSNARE Sed5 protein and stimulates its assembly into a trans-SNARE complex. Disassembly of the resulting cis-SNARE complex containing Sed5 was retarded in a temperature-sensitive yeast mutant of Slyl protein with a defect in binding to Sed5. A temperature-sensitive mutation (sec18-1) of Sec18/NSF disassembly ATPase showed synthetic lethality with the sly1(ts) mutation. These results suggest that Slyl and Sec18 proteins work cooperatively and that the binding of Slyl to Sed5 stimulates the disassembly of the cis-SNARE complex by Sec18 ATPase.

Adenosine Triphosphatases↗
PubMed2003

Effectiveness of 2-step (consecutive) embryo transfer. Comparison with cleavage-stage transfer.

OBJECTIVE: To evaluate the effectiveness of 2-step (consecutive) embryo transfer (ET), in which two cleaved embryos are transferred on day 2 (first step of ET) and a single blastocyst is transferred on day 5 (second step of ET) in comparison with conventional day 2 ET. STUDY DESIGN: Observational comparative study at different time periods. Subjects were those who had > or = 4 embryos on day 2 after oocyte retrieval. Those in the conventional group (n = 60) received conventional day 2 ET, in which 3 cleaved embryos were transferred on day 2 between January 1999 and June 2000. Those in the 2-step group (n = 124) received 2-step embryo transfer, in which 2 cleaved embryos were transferred on day 2 and a blastocyst was transferred on day 5 between September 2000 and February 2002. Patients who planned to receive 2-step ET but could not proceed to the second step of ET because of failure to produce blastocysts were included in the 2-step group. RESULTS: The ongoing pregnancy rate with 2-step ET (51.6%) was significantly higher than with conventional ET (26.7%) (P < .01). The clinical pregnancy rate and implantation rate in conventional and 2-step ET were 30.0% vs. 59.7% and 11.1% vs. 27.8%, respectively (P < .001). CONCLUSION: Two-step ET may benefit patients who can obtain a sufficient number of embryos.

Adult↗
PubMed2002

Binding of Sly1 to Sed5 enhances formation of the yeast early Golgi SNARE complex.

SLY1 is an essential gene for vesicular transport between the ER and the early Golgi apparatus in Saccharomyces cerevisiae. It encodes a hydrophilic Sec1/Munc18 family protein that binds to the t-SNAREs. The amount of Sly1 protein that coprecipitated with the t-SNARE Sed5 was much reduced in a temperature-sensitive sly1(ts) mutant yeast compared with the wildtype. The mutant Sly1(ts) protein was shown to have a reduced binding activity to Sed5. In the wildtype, a detectable amount of Sly1 was found in the complex between Sed5 and the v-SNARE Bet1. In vitro formation of this complex on different membranes in yeast lysate was enhanced by the addition of recombinant Sly1. These results indicate that binding of Sly1 to Sed5 enhances trans-SNARE complex formation.

Carrier Proteins↗
PubMed2002

A Rho GDP-dissociation inhibitor is involved in cytokinesis of Dictyostelium.

Homology searches toward the EST databases of Dictyostelium discoideum identified two putative Rho GDP-dissociation inhibitors (RhoGDIs), RhoGDI1 and RhoGDI2. In this study, the roles of RhoGDI1 in cytokinesis were examined. The RhoGDI1-null Dictyostelium strains produced by homologous recombination were viable but generated multinucleate giant cells in suspension culture, suggesting that RhoGDI1 is involved in cytokinesis. The expression of green fluorescent protein (GFP)-tagged RhoGDI1 complemented the defects of the RhoGDI1-null cells, and the GFP-RhoGDI1 is predominantly present in cytoplasm of the cell-like yeast RhoGDI. Of 15 Rho family GTPases in Dictyostelium currently known, Dictyostelium versions of Rac1 proteins (Rac1A, Rac1B, and Rac1C) and RacE that are reportedly involved in Dictyostelium cytokinesis, showed two-hybrid interactions with RhoGDI1 as well as human and yeast Cdc42. These results suggest that RhoGDI1 is involved in cytokinesis of Dicytostelium through the regulation of Rho family GTPases Rac1s and/or RacE.

Amino Acid Motifs↗
PubMed2002

Progression of the stacked Golgi compartments in the yeast Saccharomyces cerevisiae by overproduction of GDP-mannose transporter.

The Golgi compartments of the yeast Saccharomyces cerevisiaeare dispersed within the cytoplasm, in contrast to the stacked cisternae in the mammalian cell, and consequently are observed as a punctate pattern by immunofluorescent staining of Golgi-marker proteins. The VIG4/VRG4 gene encodes the essential yeast GDP-mannose transporter, which is a polytopic membrane protein in the early and medial Golgi compartments. Upon overexpression of this gene by the aid of a strong promoter and multicopy vector, we found that stacked multivesicular structures, which resembled the cisternae of mammalian Golgi apparatus, had developed in S. cerevisiae. Immuno-electron microscopy showed that the GDP-mannose transporter was located on the stacked cisternae. Immuno-isolation and immunoblotting analyses of the vesicles showed that the overproduced GDP-mannose transporter also co-localized with the Golgi glycosyltransferases, but not with the ER- or late Golgi-marker proteins as in the control cell. We propose that the localization mechanism of the GDP-mannose transporter in the Golgi compartment would be efficient and hardly saturable, and therefore the overproduced protein induced a progression of Golgi-like compartments rather than being mislocalized in other compartments, such as the ER or a vacuole.

Biological Transport↗
PubMed2002

Protein S gene mutation in a young woman with type III protein S deficiency and venous thrombosis during pregnancy.

BACKGROUND: We attempted to identify a gene defect in a young woman with type III protein S deficiency and venous thrombosis during pregnancy. METHODS: Measurements of total and free PS antigen levels in plasma were carried out using an enzyme-linked immunosorbent assay. Plasma PS cofactor activity was determined by a clotting assay using activated factor V as the substrate. Genomic DNA prepared from peripheral blood was amplified by polymerase chain reaction (PCR) with PROS1-specific oligonucleotide primers. PCR products were sequenced on both strands using specific oligonucleotide primers. RESULTS: Plasma PS cofactor activity was undetectable in every measurement at 36 weeks of gestation, as well as at 2 weeks and 4 months after delivery. Plasma total PS antigen levels were 70% and 67% at 2 weeks and 4 months after delivery, respectively. Free PS antigen level was 24% at 4 months after delivery. Of all exons analyzed, codon 295 of GGC in exon 10 was substituted for AGC. This missense mutation predicted an amino acid change of glycine to serine. CONCLUSIONS: Measurements of total and free PS antigen levels along with PS activity indicated that this was a case of type III PS deficiency. DNA analysis identified a heterozygous missense mutation of codon 295 in the PS gene, substituting glycine for serine.

Adult↗