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J Inazawa

Publications and source records attributed to J Inazawa.

At least 145 records · Page 8Linked to original sources
PubMed1996

Isolation and mapping of a human gene (DIFF6) homologous to yeast CDC3, CDC10, CDC11, and CDC12, and mouse Diff6.

We have isolated a novel human cDNA that encodes a protein homologous to murine H5 and Diff6, and to yeast CDC10, and mapped it to chromosome region 2q37 by fluorescent in situ hybridization. Its transcript has an open reading frame of 1,218 nucleotides encoding 406 amino acids. The deduced peptide sequence contained conserved domains rich in basic residues, GXXGXGKS--DXXG--TKXD, a motif of the GTPase superfamily. Different polyA sites accounted for generation of two transcripts. The major type, 3.5 kb long, was expressed ubiquitously in all human tissues examined, but a 2.0-kb alternative transcript lacking any long AU-rich element in the 3' non-coding region was expressed abundantly only in testis, heart and skeletal muscle.

Adult↗
PubMed1996

Molecular cloning of a human cDNA encoding putative cysteine protease (PRSC1) and its chromosome assignment to 14q32.1.

We have isolated a novel human cDNA encoding a protein of 433 amino acids which shows 40% sequence identity to a hemoglobinase of Schistosoma japonicum, one of the cysteine proteases in the pathway by which trematodes degrade host-cell globin. It also has 36% identity to a cysteine protease of the soybean Glycine max that processes proproteins of various vacuolar proteins into mature form after translation. Considering the sequence similarities among these proteins in such widely divergent species, the human cDNA reported here appears to represent one of a group of genes that have been well conserved during evolution. Northern-blot analysis revealed the strongest expression of this gene in kidney, among the human tissues examined. By fluorescence in situ hybridization, we determined its chromosome location at 14q32.1.

Animals↗
PubMed1996

FISH localization of human cytoplasmic actin genes ACTB to 7p22 and ACTG1 to 17q25 and characterization of related pseudogenes.

Human beta- and gamma-cytoplasmic actin genes (ACTB and ACTG1) were mapped to chromosomes 7p22 and 17q25, respectively, by fluorescence in situ hybridization (FISH). Four processed pseudogenes, beta-actin-related ACTBP9 and gamma-actin-related ACTGP1, ACTGP3, and ACTGP9, were isolated from human libraries. By PCR of somatic cell hybrid DNAs, ACTBP9 and two beta-actin-related pseudogenes (ACTBP7 and ACTBP8) were mapped to human chromosomes 18, 15, and 6, respectively. The gamma-actin-related pseudogenes were mapped by FISH to chromosomes 3q23 (ACTGP1), 20p13 (ACTGP3), and 6p21.1 (ACTGP9).

Actins↗
PubMed1996

Molecular cloning and mapping of a human cDNA (SC5DL) encoding a protein homologous to fungal sterol-C5-desaturase.

We have isolated a human cDNA clone homologous to fungal ERG3, a gene encoding sterol C-5 desaturase. The full nucleotide sequence of this human cDNA revealed a 708-bp open reading frame that encodes a 236-amino-acid polypeptide. The gene was expressed in all normal human tissues examined. We determined its location to chromosome 11q23.3 by fluorescence in situ hybridization.

Amino Acid Sequence↗
PubMed1996

Induction of mcl1/EAT, Bcl-2 related gene, by retinoic acid or heat shock in the human embryonal carcinoma cells, NCR-G3.

NCR-G3 cells were established from a testicular embryonal carcinoma and were differentiated into multi-lineages including trophectoderm cells by exposure to retinoic acid. The differentiated cells began to produce human chorionic gonadotropin (hCG), a trophectoderm-specific hormone, which was regulated at the mRNA level. As we assumed that genes responsible for differentiation were differentially expressed at the early stage of retinoic acid-induced differentiation, we prepared a cDNA library from retinoic acid-treated NCR-G3 cells. This cDNA library was then screened for genes whose expression was induced during the differentiation of these cells. From about 5 x 10(4) clones screened, three independent sequences were isolated. Sequencing analysis revealed that clone 1002 codes for mcl1/EAT, which has a Bcl-2 homology domain. The expression of mcl1/EAT, the Bcl-2 related gene, was increased at an early stage of the retinoic acid-induced differentiation and preceded the up-regulation of cytokeratin and hCG genes after ratinoic acid treatment. Furthermore, mcl1/EAT was also up-regulated by heat shock, which has recently been shown to induce the cells to differentiate.

Biomarkers↗
PubMed1996

Abnormal expression of Evi-1 gene in human leukemias.

The human Evi-1 gene located on chromosome 3q26, encodes a zinc finger protein that functions as a transcription factor. It was frequently overexpressed in leukemias having 3q26 abnormalities such as t(3;3)(q21;q26) and inv(3)(q21 q26), and subjected to structural alteration in t(3;21)(q26;q22). In addition, recent studies indicated that several cases of leukemias without 3q26 abnormalities also expressed Evi-1 gene. In this study we present another case of structural alteration of Evi-1 gene in a case of inv(3)(q21 q26), in which Evi-1 was truncated and a shorter form of Evi-1 protein was expressed upon rearrangement of the gene. We also studied expression of the Evi-1 gene in a variety of leukemias by northern blot analysis. Evi-1 was overexpressed not only in leukemias with 3q26 abnormalities, but, in those without 3q26 abnormalities, especially in blast crisis of CML. Our result also supports an idea that Evi-1 is a relevant oncogene whose overexpression or structural changes might play a crucial role in development of human leukemias.

Adult↗
PubMed1995

Molecular cloning and chromosomal assignment of the human brain-type phosphodiesterase I/nucleotide pyrophosphatase gene (PDNP2).

Phosphodiesterase I/nucleotide pyrophosphatase is a widely expressed membrane-bound enzyme that cleaves diester bonds of a variety of substrates. We have cloned brain-type cDNA for this enzyme from rat brain and designated it PD-I alpha (M. Narita, J. Goji, H. Nakamura, and K. Sano, 1994, J. Biol. Chem. 269: 28235-28242). In this study we have isolated cDNA and genomic DNA encoding human PD-I alpha. Human PD-I alpha cDNA, designated PDNP2 in HGMW nomenclature, has a 2589-nucleotide open reading frame encoding a polypeptide of 863 amino acids with a calculated M(r) of 99,034. Northern blot analysis revealed that human PD-I alpha transcript was present in brain, lung, placenta, and kidney. The database analysis showed that human PD-I alpha was identical with human autotaxin (ATX), a novel tumor motility-stimulating factor, except that human PD-I alpha lacks 156 nucleotides and 52 amino acids of human ATX. Human PD-I alpha and human ATX are likely to be alternative splicing products from the same gene. The 5' region of the human PDNP2 gene contains four putative binding sites of transcription factor Sp1 without typical TATA or CAAT boxes, and there is a potential octamer binding motif in intron 2. From the results of fluorescence in situ hybridization, the human PDNP2 gene is located at chromosome 8q24.1.

Base Sequence↗
PubMed1995

Isolation and mapping of human homologues of an imprinted mouse gene U2af1-rs1.

We have isolated human homologues of the imprinted mouse gene, U2af1-rs1. Two different types of cDNAs and three distinct genomic DNAs belonging to different groups were isolated. We have identified chromosomal genes corresponding to each cDNA by restriction mapping and sequencing. Using both a panel of rodent/human somatic cell hybrids and fluorescence in situ hybridization, group 1 and group 2 genes were mapped to chromosome 5q22 and chromosome Xp22.1, respectively. We designated group 1 and group 2 genes as human U2AF1-RS1 and U2AF1-RS2, respectively, because these genes corresponded to mouse U2af1-rs1 (chromosome 11) and U2af1-rs2 (chromosome X), which we also isolated and mapped. Amino acid sequences of human U2AF1-RS1 and U2AF-RS2 showed significant homology to U2AF small subunit. The group 3 gene, designated as U2AF1-RS3, of which the cDNA has not yet been isolated, was mapped to chromosome 19p13.2.

Amino Acid Sequence↗
PubMed1995

Reconstitution of IKATP: an inward rectifier subunit plus the sulfonylurea receptor.

A member of the inwardly rectifying potassium channel family was cloned here. The channel, called BIR (Kir6.2), was expressed in large amounts in rat pancreatic islets and glucose-responsive insulin-secreting cell lines. Coexpression with the sulfonylurea receptor SUR reconstituted an inwardly rectifying potassium conductance of 76 picosiemens that was sensitive to adenosine triphosphate (ATP) (IKATP) and was inhibited by sulfonylureas and activated by diazoxide. The data indicate that these pancreatic beta cell potassium channels are a complex composed of at least two subunits--BIR, a member of the inward rectifier potassium channel family, and SUR, a member of the ATP-binding cassette superfamily. Gene mapping data show that these two potassium channel subunit genes are clustered on human chromosome 11 at position 11p15.1.

ATP-Binding Cassette Transporters↗
PubMed1995

cDNA sequence, gene structure, and chromosomal localization of the human ATP-sensitive potassium channel, uKATP-1, gene (KCNJ8).

ATP-sensitive K+ (KATP) channels play a crucial role in coupling metabolic energy to the membrane potential of cells. Recently, we have isolated a KATP channel cDNA (uKATP-1) that is expressed ubiquitously in rat tissues including pancreatic islets, pituitary, skeletal muscle, and heart. Here, we report cloning of the human cDNA and gene encoding uKATP-1. Human uKATP-1 is a protein of 424 amino acids exhibiting 98% identity with rat uKATP-1. The human gene encoding uKATP-1, designated KCNJ8, is approximately 9.7 kb in length and is composed of three exons. KCNJ8 was mapped to chromosome 12p11.23 using fluorescence in situ hybridization.

ATP-Binding Cassette Transporters↗
PubMed1995

The human protoporphyrinogen oxidase gene (PPOX): organization and location to chromosome 1.

We determined the structure of the human protoporphyrinogen oxidase (PPOX) gene after isolation and characterization of lambda phage clones mapping discrete regions of the cDNA. Southern blotting of human genomic DNA showed that there is a single copy of the PPOX gene, and fluorescence in situ hybridization to metaphase chromosomes mapped the gene to region 1q22. The gene has 13 exons and about 8 kb. The exon/intron boundary sequences conform to consensus acceptor (GTn) and donor (nAG) sequences, and exons in the gene appear to encode functional protein domains. Primer extension analysis revealed two major transcriptional initiation sites in a region with sequence motifs characteristic of a promoter. The promoter region contains multiple Sp1 elements, CCAAT boxes, and potential GATA-1 binding sites. Mapping of the 5' end PPOX mRNA by polymerase chain reaction indicated that there are the same transcripts in erythroid and nonerythroid cells.

Bacteriophage lambda↗
PubMed1995

ERM, a PEA3 subfamily of Ets transcription factors, can cooperate with c-Jun.

A human cDNA clone for ERM, a member of the ets gene family, has been obtained by polymerase chain reaction with degenerate primers corresponding to highly conserved regions within an Ets DNA binding domain. ERM mRNA is expressed ubiquitously. The gene was mapped to chromosome 3q27. In in vivo transient-expression assays, ERM induced transcription more efficiently from a synthetic element containing both an ets-binding site (EBS) and a cyclic AMP response element (CRE) than from one containing an EBS alone. The activation of a synthetic EBS-CRE site by ERM was likely to involve a leucine zipper protein capable of dimerizing with CRE-BP1 leucine zipper. Indeed, ERM and c-Jun synergistically activated the EBS-CRE without making an apparent ternary complex. The synergy between c-Jun and ERM may be attributed to the enhancing effect of c-Jun on the transcription activity of ERM, because c-Jun increased ERM transcription activity by more than 20-fold in an assay system using a variety of fusion proteins between a Gal4 DNA-binding domain and a portion of ERM. This enhancing effect of c-Jun required the amino-terminal portion of ERM.

3T3 Cells↗
PubMed1995

Structure and chromosomal localization of the human stromal cell-derived factor 1 (SDF1) gene.

Stromal cell-derived factors 1 alpha and 1 beta are small cytokines belonging to the intercrine CXC subfamily and originally isolated from a murine bone-marrow stroma cell line by the signal sequence trap method. cDNA and genomic clones of human SDF1 alpha and SDF1 beta (SDF1A and SDF1B) were isolated and characterized. cDNAs of SDF1 alpha and SDF1 beta encode proteins of 89 and 93 amino acids, respectively. SDF1 alpha and SDF1 beta sequences are more than 92% identical to those of the human counterparts. The genomic structure of the SDF1 gene revealed that human SDF1 alpha and SDF1 beta are encoded by a single gene and arise by alternative splicing. SDF1 alpha and SDF1 beta are encoded by 3 and 4 exons, respectively. Ubiquitous expression of the SDF1 gene, except in blood cells, was consistent with the presence of the GC-rich sequence in the 5'-flanking region of the SDF1 gene, as is often the case in the "housekeeping" genes. Although genes encoding other members of the intercrine family are localized on chromosome 4q or 17q, the human SDF1 gene was mapped to chromosome 10q by fluorescence in situ hybridization. Strong evolutionary conservation and unique chromosomal localization of the SDF1 gene suggest that SDF1 alpha and SDF1 beta may have important functions distinct from those of other members of the intercrine family.

Amino Acid Sequence↗
PubMed1995

Chromosomal loci of 50 human keratinocyte cDNAs assigned by fluorescence in situ hybridization.

The chromosomal loci of expressed genes provide useful information for a candidate gene approach to the genes responsible for genetic diseases. A large set of randomly isolated cDNAs catalogued by partial sequencing can serve as a resource for accessing and isolating these disease genes. Using fluorescence in situ hybridization, we examined the chromosomal loci of 217 human keratinocyte-derived cDNAs, with independent novel sequence tags at the 3' end region. Among them, we determined the loci of 50 cDNAs. Single-pass sequencing of these from the 5' ends indicated that 39 cDNAs still can be produced for new genes. These cDNAs with identified chromosomal loci are powerful tools that can be used to help elucidate the genes responsible for hereditary skin disorders.

Chromosome Mapping↗
PubMed1995

Elevated expression of Unph, a proto-oncogene at 3p21.3, in human lung tumors.

The murine Unp proto-oncogene encodes a nuclear ubiquitin protease whose overexpression leads to oncogenic transformation of NIH3T3 cells. We have cloned cDNAs originating from the human homolog of this gene, designated herein as Unph (Unp, human), and have used these cDNAs to map the gene to 3p21.3, a region frequently rearranged in human tumor cells. Unph mRNA levels are consistently elevated in small cell tumors and adenocarcinomas of the lung, suggesting a possible causative role for the gene in the neoplastic process.

3T3 Cells↗