Biomedical subjects

N Minato

Publications and source records attributed to N Minato.

At least 37 records · Page 2Linked to original sources

PubMed1999

Identification of the von Hippel-lindau tumor-suppressor protein as part of an active E3 ubiquitin ligase complex.

Mutations of von Hippel-Lindau disease (VHL) tumor-suppressor gene product (pVHL) are found in patients with dominant inherited VHL syndrome and in the vast majority of sporadic clear cell renal carcinomas. The function of the pVHL protein has not been clarified. pVHL has been shown to form a complex with elongin B and elongin C (VBC) and with cullin (CUL)-2. In light of the structural analogy of VBC-CUL-2 to SKP1-CUL-1-F-box ubiquitin ligases, the ubiquitin ligase activity of VBC-CUL-2 was examined in this study. We show that VBC-CUL-2 exhibits ubiquitin ligase activity, and we identified UbcH5a, b, and c, but not CDC34, as the ubiquitin-conjugating enzymes of the VBC-CUL-2 ubiquitin ligase. The protein Rbx1/ROC1 enhances ligase activity of VBC-CUL-2 as it does in the SKP1-CUL-1-F-box protein ligase complex. We also found that pVHL associates with two proteins, p100 and p220, which migrate at a similar molecular weight as two major bands in the ubiquitination assay. Furthermore, naturally occurring pVHL missense mutations, including mutants capable of forming a complex with elongin B-elongin C-CUL-2, fail to associate with p100 and p220 and cannot exhibit the E3 ligase activity. These results suggest that pVHL might be the substrate recognition subunit of the VBC-CUL-2 E3 ligase. This is also, to our knowledge, the first example of a human tumor-suppressor protein being directly involved in the ubiquitin conjugation system which leads to the targeted degradation of substrate proteins.

Carrier Proteins↗

PubMed1999

Rap1 GTPase-activating protein SPA-1 negatively regulates cell adhesion.

Rap1 GTPase is activated by a variety of stimulations in many types of cells, but its exact functions remain unknown. In this study we have shown that SPA-1 interferes with Rap1 activation by membrane-targeted C3G, C3G-F, in 293T cells through the GTPase activating protein (GAP) activity. SPA-1 transiently expressed in HeLa cells was mostly localized at the cortical cytoskeleton and induced rounding up of the cells, whereas C3G-F conversely induced extensive cell spreading. Conditional SPA-1 overexpression in HeLa cells by tetracycline-regulative system suppressed Rap1 activation upon plating on dishes coated with fibronectin and resulted in the reduced adhesion. When SPA-1 was conditionally induced after the established cell adhesion, the cells gradually rounded up and detached from the dish. Both effects were counteracted by exogenous fibronectin in a dose-dependent manner. Retroviral overexpression of SPA-1 in promyelocytic 32D cells also inhibited both activation of Rap1 and induction of cell adhesion by granulocyte colony stimulating factor without affecting differentiation. These results have indicated that Rap1 GTP is required for the cell adhesion induced by both extracellular matrix and soluble factors, which is negatively regulated by SPA-1.

Animals↗

PubMed1999

The bHLH gene Hes1 is essential for expansion of early T cell precursors.

Mice mutant for the bHLH gene Hes1, which is known to keep cells in a proliferative state, mostly lack thymus. Transfer of Hes1-null fetal liver cells into RAG2-null host mice normally reconstitutes B cells but fails to generate mature T cells in the thymus. In the reconstituted thymus, T cell differentiation is arrested at the CD4(-)CD8(-) double negative (DN) stage. Both the initial T cell receptor (TCR)-independent and the subsequent TCR-dependent selective expansion during the DN stage are severely affected. Thus, Hes1 is essential for the earliest thymocyte expansion in a cell-autonomous manner.

Animals↗

PubMed1999

4F2 (CD98) heavy chain is associated covalently with an amino acid transporter and controls intracellular trafficking and membrane topology of 4F2 heterodimer.

4F2, also termed CD98, is an integral membrane protein consisting of a heavy chain linked to a light chain by disulfide bond. We have generated a monoclonal antibody to the mouse 4F2 light chain and cloned the cDNA. It encodes a mouse counterpart of rat L-type amino acid transporter-1, and induces system L amino acid transport in Xenopus oocytes in the presence of 4F2 heavy chain. Transfection studies in mammalian cells have indicated that the 4F2 heavy chain is expressed on the plasma membrane on its own, whereas the 4F2 light chain can be transported to the surface only in the presence of 4F2 heavy chain. 4F2 heavy chain is expressed diffusely on the surface of fibroblastic L cells, whereas it is localized selectively to the cell-cell adhesion sites in L cells expressing cadherins. These results indicate that the 4F2 heavy chain is associated covalently with an amino acid transporter and controls the cell surface expression as well as the membrane topology of the 4F2 heterodimer. Although 4F2 heavy and light chains are expressed coordinately in most tissues, the light chain is barely detected by the antibody in kidney and intestine, despite the presence of heavy chain in a complex form. The results predict the presence of multiple 4F2 light chains.

Amino Acid Transport Systems↗

PubMed1999

Development of lupus-like autoimmune diseases by disruption of the PD-1 gene encoding an ITIM motif-carrying immunoreceptor.

PD-1, a 55 kDa transmembrane protein containing an immunoreceptor tyrosine-based inhibitory motif, is induced in lymphocytes and monocytic cells following activation. Aged C57BL/6(B6)-PD-1(-/-) congenic mice spontaneously developed characteristic lupus-like proliferative arthritis and glomerulonephritis with predominant IgG3 deposition, which were markedly accelerated by introduction of a Fas mutation (lpr). Introduction of a PD-1 null mutation into the 2C-TCR (anti-H-2Ld) transgenic mice of the H-2(b/d) background resulted in the chronic and systemic graft-versus-host-like disease. Furthermore, CD8+ 2C-TCR+ PD-1(-/-) T cells exhibited markedly augmented proliferation in vitro in response to H-2d allogenic cells. Collectively, it is suggested that PD-1 is involved in the maintenance of peripheral self-tolerance by serving as a negative regulator of immune responses.

Animals↗

PubMed1999

Surgery of the dissecting aneurysm involving a right aortic arch.

A dissecting aneurysm in association with a right aortic arch is extremely rare. However, a 50-year-old male was diagnosed as having a dissecting aneurysm (DeBakey IIIa) with a right aortic arch, right descending aorta and an aberrant retro-esophageal left subclavian artery. A graft replacement of the right descending aorta was successfully performed under right thoracotomy and partial cardiopulmonary bypass. Precise anatomical definition and proper surgical procedure permitted a successful surgical result.

Aortic Dissection↗

PubMed1999

Mitral valve replacement through a giant left atrial appendage.

We report a rare case of a 45-year-old male with a giant left atrial appendage (13x10 cm in size) and chronic massive regurgitation at the mitral valve. Massive dilatation was compressing most of the left lower lobe and the large size of the appendage was pushing the mediastinum to the right. Therefore under median sternotomy, the mitral valve was clearly accessible and chordal sparing mitral valve replacement with left atrial plication was successfully performed through the giant left atrial appendage.

Adult↗

PubMed1999

[Emergency coronary artery bypass grafting for severe ischemia in patients with left main trunk disease].

Between Jan. 1994 and Feb. 1998, thirty-eight patients underwent emergency coronary artery bypass grafting (CABG) for severe left main trunk (LMT) disease. The age ranged from 45 to 87 (mean 69.8), including 10 patients over 80 years. Surgical indications were: acute myocardial infarct (AMI) due to acute occlusion of LMT (LMT-AMI) in 7, unstable angina with severe LMT stenosis (LMT-UAP) in 19, AMI due to branch occlusion with severe LMT stenosis (AMI with LMT) in 10, and chronic LMT occlusion with ongoing congestive heart failure (LMT-OMI) in 2 patients. Five of the 7 patients with LMT-AMI were successfully resuscitated from ventricular fibrillation at the emergency room in 2 (cardio-pulmonary arrest on arrival), and at the cathe-room in 3 patients. For these patients, percutaneous cardiopulmonary support system + intra aortic balloon pump (IABP) were required in 3, and IABP in 2 patients before or during catheter intervention. LMT was then recanalized by catheter intervention prior to CABG. Operative mortality were 28.6% (2/7) in LMT-AMI, 20% (2/10) in AMI with LMT, 0% (0/19) in LMT-UAP, and 50% (1/2) in LMT-OMI. Five of the 7 patients with LMT-AMI, survived the most hazardous condition of left main shock syndrome. Quick establishment of the circulatory support and recanalization of LMT by catheter intervention prior to CABG are the key factors for saving their lives.

Aged↗

PubMed1999

Does dilatation of the sinotubular junction cause aortic regurgitation?

BACKGROUND: Some patients develop aortic regurgitation (AR) in association with dilatation of the sinotubular junction (STJ), despite having normal aortic valve. However, the relationship between dilatation of the STJ and AR is unclear. METHODS: Canine hearts and aortas were isolated. A suture was placed in each commissure and in the sinus of Valsalva at the STJ. These interrupted sutures were drawn horizontally, and strain on the sutures was varied. The sites of the retracted sutures were changed to various positions, and the opening and closing of the aortic valve was observed endoscopically. A beating heart model was used to observe changes in aortic valve function during mechanical retraction of the commissures or sinuses. RESULTS: Opening area of the valve increased when strain on all sutures or commissures was increased. When strain was increased on the sinus alone, coaptation of the valve was not affected. CONCLUSION: We observed endoscopically that mechanical dilatation of the STJ causes AR. These findings suggest that the principal cause of AR associated with dilatation of the STJ is outward deviation of the commissure.

Animals↗

PubMed1999

Invasive thymoma with intracaval growth extending and directly invading the right atrium.

We report an extremely rare case of an invasive thymoma with intracaval growth extending and directly invading the right atrium. Enbloc excision of the tumor with lobectomy and combined resection of the right atrium, and graft replacement of the superior vena cava were successfully performed during cardiopulmonary bypass. Invasive thymoma with this type of intracaval and intra-atrial progression is extremely rare.

Adult↗

PubMed1998

Iron-dependent oxidation, ubiquitination, and degradation of iron regulatory protein 2: implications for degradation of oxidized proteins.

The ability of iron to catalyze formation of reactive oxygen species significantly contributes to its toxicity in cells and animals. Iron uptake and distribution is regulated tightly in mammalian cells, in part by iron regulatory protein 2 (IRP2), a protein that is degraded efficiently by the proteasome in iron-replete cells. Here, we demonstrate that IRP2 is oxidized and ubiquitinated in cells before degradation. Moreover, iron-dependent oxidation converts IRP2 into a substrate for ubiquitination in vitro. A regulatory pathway is described in which excess iron is sensed by its ability to catalyze site-specific oxidations in IRP2, oxidized IRP2 is ubiquitinated, and ubiquitinated IRP2 subsequently is degraded by the proteasome. Selective targeting and removal of oxidatively modified proteins may contribute to the turnover of many proteins that are degraded by the proteasome.

Animals↗

PubMed1998

[A case of the ascending aorta and aortic arch replacement with thrombo-occlusion of distal arch and descending aorta surround the modified elephant trunk graft].

We report a 65-year-old female who had a extensive thoracic aneurysm from ascending aorta to descending thoracic aorta. The patient underwent a graft replacement of ascending aorta and aortic arch using modified elephant trunk method. The surgery was carried out through median sternotomy with profound hypothermia and selective cerebral perfusion. Postoperatively, the patient was recovered without any complications except bronchial asthma. Postoperative chest computed tomography showed that the surrounding space of the elephant trunk vascular graft inserted into distal arch and descending aneurysm was mostly occupied with thrombus. Therefore, we considered that the second operation on the descending aorta is not necessary at this point and careful attention to the size and shape of the descending aneurysm should be paid.

Aged↗

PubMed1998

Immunological studies on PD-1 deficient mice: implication of PD-1 as a negative regulator for B cell responses.

PD-1, an Ig superfamily member, contains an immunoreceptor tyrosine-based inhibitory motif in the cytoplasmic tail. It is expressed in a minor fraction of CD4-CD8- normal thymocytes and induced in peripheral lymphocytes following activation. To assess the possible roles of PD-1 in the immune responses, PD-1-deficient (PD-1-/-) mice were generated by a gene-targeting strategy. PD-1-4- mice developed and grew normally. Although the thymus was apparently normal, PD-1-/- mice showed moderate but consistent splenomegaly, which reflected the increased cellularity of both lymphoid and myeloid cells. The proliferative response of B cells by anti-IgM antibodies, but not of T cells by an anti-CD3 (145-2C11) mAb in vitro, was augmented in PD-1-/- mice as compared with control littermates. PD-1-/- mice showed increased serum levels of IgG2b, IgA and most strikingly IgG3, while those of IgM and IgG1 were comparable with control mice. Furthermore, PD-1-/- mice exhibited significantly augmented IgG3 anti-DNP antibody response to a type 2 T-independent antigen, DNP-Ficoll, with comparable IgM and IgG1 antibody responses with littermate controls. In the peritoneal cavity, the B-1 cell population in PD-1-/- mice exhibited significantly reduced expression of CD5, a negative regulator of B-1 cell activation, despite a marginal increase in the number of B-1 cells. Thus, PD-1 was suggested to be involved in the negative regulation for particular aspects of B cell proliferation and differentiation including class switching.

Animals↗

PubMed1998

Mutational analysis of superantigen activity responsible for the induction of skin erythema by streptococcal pyrogenic exotoxin C.

Streptococcal pyrogenic exotoxin C (SPEC), when injected intradermally, induces erythema in unsensitized rabbits. In the present study, we examined whether this erythema induction is due to the T-cell stimulatory activity of SPEC as a superantigen. Analysis by using single-residue mutant SPECs indicated that mutant SPECs Y15I, A16E, and Y17I, in which tyrosine 15, alanine 16, and tyrosine 17 were replaced with isoleucine, glutamic acid, and isoleucine, respectively, exhibited significantly reduced mitogenic activity for Vbeta2(+) human T cells in vitro, and Y15I showed as much as a 1, 000-fold reduction. Y15I mutant SPEC, however, retained the ability to bind to major histocompatibility complex class II antigen and to form a homodimer, implying that residue 15 is critically important for the interaction of SPEC with T-cell antigen receptor beta chains. When injected intradermally into normal rabbits, wild-type SPEC induced a characteristic erythema after 3 h in a dose-dependent fashion, which was associated with polymorphonuclear and mononuclear cell infiltration. This erythema formation was found to be severely suppressed by systemic pretreatment with cyclosporin A, suggesting the involvement of host T cells. Y15I mutant SPEC exhibited nearly 1, 000-fold less erythema induction in vivo than wild-type SPEC. Altogether, the present results strongly suggest that erythema induction in rabbits by SPEC is attributable mostly to its T-cell stimulatory activity as a superantigen.

Amino Acid Sequence↗

PubMed1997

Human SPA-1 gene product selectively expressed in lymphoid tissues is a specific GTPase-activating protein for Rap1 and Rap2. Segregate expression profiles from a rap1GAP gene product.

Mouse Spa-1 gene with a region homologous to the human rap1GAP gene is transcriptionally induced in the lymphocytes by mitogenic stimulation. Herein we have cloned a cDNA for its human counterpart. SPA-1 cDNA encodes a 130-kDa protein (p130(SPA-1)) consisting of proline-rich regions and rap1GAP-related domain followed by a coiled-coil stretch. Baculovirally expressed p130(SPA-1) exhibited GTPase-activating protein (GAP) activity for Rap1 and Rap2, but not for Ras, Rho, Cdc42, Rac, and Ran, with comparable specific activity to the rap1GAP gene product (p85/95(rap1GAP)). In the cells, p130(SPA-1) was mostly localized at the perinuclear membranous region co-localizing with Rap1 and Rap2. Expression of SPA-1 and rap1GAP genes tended to be segregate in various tissues, lymphoid tissues expressing abundant SPA-1 transcript without rap1GAP, while those such as brain, kidney, and pancreas exhibiting rap1GAP mRNA with little SPA-1. Promyelocytic HL-60 cells, which expressed p130(SPA-1) with little p85/95(rap1GAP) in uninduced state, showed progressive decline in p130(SPA-1) and conversely drastic increase in p85/95(rap1GAP) as they ceased from proliferation and differentiated into macrophages by 12-O-tetradecanoylphorbol-13-acetate. These results suggested that products of SPA-1 and rap1GAP genes, albeit comparable GAP activity for Rap1 and Rap2, functioned in the distinct contexts depending on cell types and/or states.

Amino Acid Sequence↗

PubMed1997

Involvement of NK1+ CD4- CD8- alphabeta T cells and endogenous IL-4 in non-MHC-restricted rejection of embryonal carcinoma in genetically resistant mice.

Non-MHC-restricted rejection mechanisms against the murine MHC-negative F9 embryonal carcinoma cells were analyzed. Strains of C57BL/6 (B6) background were resistant to the tumors irrespective of H-2 haplotypes, while others, including BALB/c background, were susceptible. This resistance was suggested to be mediated primarily by the host thymus-dependent alphabeta T cells, since both athymic B6 nude and normal B6 mice depleted with alphabeta T cells showed susceptible phenotype. The difference of the nature of alphabeta T cells infiltrating in H-2-identical B6- and BALB.B-derived tumors was then comparatively analyzed. It was revealed that unique T cells with NK1+ CD4- CD8- (double negative (DN)) alphabeta TCR+ phenotype were accumulated significantly in B6, but few in BALB.B mice. The population freshly isolated from the F9 tumor tissues preferentially expressed potent IL-4 mRNA, and was suggested to be mostly responsible for the endogenous IL-4 production. Indeed, the injection of either anti-NK1.1 or anti-IL-4-neutralizing Ab into the normal B6 rendered them significantly susceptible to the tumor cells. These results strongly suggested that NK1+ DN alphabeta T cells were responsible primarily for the rejection mechanisms against F9 tumors. Histologically, F9 tumors in B6 mice were characterized by abundant macrophage infiltration and massive tumor necrosis, neither of which was observed in those in BALB.B nor B6 mice preinjected with anti-IL-4 Ab, indicating that both histologic features in the resistant strain were dependent on the endogenous IL-4. Present results provide one of the first instances in which a recently emerging minor T cell subpopulation, thymus-dependent NK1+ DN alphabeta T cells, plays an essential role in anti-tumor responses in vivo.

Animals↗

PubMed1997

Mitogen-inducible SIPA1 is mapped to the conserved syntenic groups of chromosome 19 in mouse and chromosome 11q13.3 centromeric to BCL1 in human.

Sipa1, previously called Spa1, is transcriptionally induced in the murine lymphoid cells following mitogenic stimulation and encodes a protein with a domain related to Rap1 GTPase activating protein (Rap1GAP) at the N-terminus and to PEST sequences followed by a leucine zipper motif at the C-terminus. Herein mouse genomic Sipa1, which consisted of 16 exons, was cloned. Gene linkage analysis using (BXD) recombinant inbred strains indicated that Sipa1 was mapped to the most centromeric region of chromosome 19 syntenic with the long arm of human chromosome 11. Human SIPA1 cDNA exhibited a striking homology to that of mouse throughout the entire region, with the overall identity being 90% at the amino acid level. Human genomic clones, which hybridized with both mouse and human SIPA1 cDNA but not with RAP1GAP cDNA, were then isolated. Fluorescence in situ hybridization (FISH) analysis using the human genomic clones indicated that SIPA1 was indeed mapped to chromosome 11q13, most likely to the 11q13.3 subregion. It was further indicated by double-color FISH that SIPA1 was located in the centromeric neighborhood of CCND1/ PRAD1, a presumed BCL1 oncogene.

Animals↗

PubMed1997

[A successful surgical case report of impending rupture of the true thoraco-abdominal aortic aneurysm induced by thrombosed-type acute aortic dissection (Stanford type A)].

Impending rupture of the true aneurysm of the thoraco-abdominal aorta induced by acute aortic dissection (Stanford type A) is very rare, and decision making of the therapeutic plain is difficult. A 88-year-old woman manifested severe back pain with hypotension. Chest computed tomographic examination revealed a true aneurysm of the thoraco-abdominal aorta and acute thrombosed-type aortic dissection (Stanford type A) with cardiac tamponade. Under the diagnosis of impending rupture of the thoraco-abdominal aortic aneurysm induced by acute aortic dissection (Stanford type A), graft replacement of the thoraco-abdominal aortic aneurysm and pericardial drainage were successfully performed. Two months later, computed tomographic examination revealed disappearance of the thrombosed false lumen in the ascending aorta.

Acute Disease↗