Biomedical subjects
Publications and source records attributed to M C Yoshida.
Pyruvate kinase (PK) is an important enzyme for ATP production in the glycolytic pathway. Deficiency of this enzyme in erythrocytes is characterized by hemolytic anemia. Using in situ hybridization, we have mapped the human liver-type pyruvate kinase gene (PKL) to band q21 of chromosome 1.
A novel human oncogene, LCA, was assigned to region 2q14----q21 by in situ molecular hybridization. The present regional mapping substantiates the previous assignment that was performed by Southern blot analyses of DNAs from flow-sorted human chromosomes and human-mouse somatic cell hybrids.
The natural history of hereditary hepatitis in long-survived LEC rats was reported. Among 56 (female: male, 28:28) LEC rats of F30, 16 (8:8) (29%) died of fulminant hepatitis approximately four months after birth. The remaining 40 (20:20) rats that survived more than one year developed chronic hepatitis and subsequent hepatic lesions including hepatocellular carcinomas. Further study made with 32 F31 rats killed at the age of five months revealed that hepatitis occurred in all of these rats. Genetic analysis performed by various crosses of LEC and LEJ rats confirmed the previous result that hereditary hepatitis was caused by a single autosomal recessive gene. F1 hybrid rats never developed hepatitis, showing normal histology of the liver. Histological features of hepatitis in F2 (F1 X F1) and backcross (F1 X LEC) rats were the same as those observed in the LEC rats. The preneoplastic foci also appeared in some of these hybrid rats at the age of eight months. We propose a gene symbol hts to designate the present hepatitis which is assumed to be homozygous in LEC strain rats.
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We previously isolated a cDNA clone for human cytochrome c1. The insert DNA of approximately 950 bp from this clone was used as a probe to identify the cytochrome c1 gene. High molecular weight DNAs extracted from a panel of 14 independent human-mouse somatic cell hybrids were digested with BamHI and analyzed by Southern blot hybridization. The results indicated that the gene for human cytochrome c1 is located on chromosome 8.
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Pyruvate kinase (PK) has four isozymes (L,R,M1,M2) that are encoded mainly by two different genes. We isolated a cDNA clone from a Japanese adult liver lambda gt10 cDNA library by using a rat liver(L)-type PK cDNA probe. One positively hybridizing clone, hlPK-1, which contained a 1,049-base pair cDNA insert, was subjected to DNA sequence analysis. Comparisons of the sequence data with the rat PK cDNAs indicated that the cDNA encoded information for the carboxyl terminal 105 amino acids of a human L-type PK and a 3' untranslated region of 734 nucleotides. Furthermore, the karyotype analysis of several human-mouse hybrid cells and Southern blot analysis of DNAs of the hybrids with a hlPK-1 indicated that the human L-type PK gene is located on chromosome 1.
Mapping of human aldolase A (ALDOA) gene was performed by molecular hybridization techniques using a panel of human-mouse cell hybrids and sorted fractions of human metaphase chromosomes besides in situ hybridization. For the purpose, three kinds of DNA probes derived from the coding region (probe-1), the 3' noncoding region (probe-2), and the coding and 3' noncoding regions (probe-3) of human aldolase A cDNA clone, pHAAL116-3, were selectively employed. The results of RNA and DNA blot analyses indicated that the human ALDOA gene is located on chromosome 16. The in situ hybridization experiment also indicated that the ALDOA gene was localized to 16q22-q24.
A human-mouse hybrid segregant HM76Dd40-6 with new characteristics was derived from the hybrid cell line HM76Dd containing human chromosome 19 as the only human chromosome. Three virus sensitivities located on human chromosome 19 (PVS, E11S and RDRC) were lost in HM76Dd40-6, while six other genes (C3, LDLR, EF2, GPI, PEPD and MANB) were retained. Cytogenetic analysis and in situ hybridization using human or mouse repeated sequences as probes showed that the region q13.1-qter of human chromosome 19 had been replaced by a fragment of mouse chromosome. Our results permit further regional assignment for the following five genes on human chromosome 19: GPI in the region cen-q12, MANB in p13.2-q12, E11S and RDRC in q13.1-qter, and EF2 in pter-q12.
Granulocyte-colony stimulating factor (G-CSF) is a member of colony stimulating factors which regulate the proliferation and differentiation of hematopoietic progenitor cells. A full-length cDNA clone coding human G-CSF was used as a hybridization probe to detect homologous sequence in human-mouse somatic cell hybrids, flow-sorted human chromosomes, and in situ human metaphase chromosomes. The results indicate that the gene encoding human G-CSF is on the q21-q22 region of chromosome 17, which is involved in translocation of t(15;17) (q23;21) in human acute promyelocytic leukemia.
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The tsBN462 cell line, a temperature-sensitive (ts) mutant isolated from the hamster cell line, BHK21/13, cannot progress into S phase at 39.5 degrees C, following the release from isoleucine deprivation. The mutant cells were transfected with high molecular weight (HMW) DNA from human KB cells, and several human DNA bands were found to be conserved through three cycles of ts+ transformation. Conserved human DNA was isolated from the cosmid library of the secondary ts+ transformant (K-1-1), using 32P-labelled total human DNA as a probe. The isolated human DNA covers about 70 kb of human DNA flanked with hamster DNA, and originates from the human X chromosome. The middle part (56 kb) of the isolated human DNA was conserved through the primary, secondary and tertiary ts+ transformation, without gross rearrangement.
DNA rearrangements that occurred in the vicinity of T-cell antigen receptor beta-chain gene clusters residing on chromosome 7 were examined in human T-cell acute lymphoblastic leukemia cells. In one patient, it was observed that, for the T-cell receptor beta-chain genes, a D beta 1-J beta 2.3 (where D is diversity and J is joining) junction was found on one chromosome, while the other chromosome kept the germ-line configuration. If this D beta-J beta junction was formed by the customary deletion mechanism, the C beta 1 gene (where C is constant) located between the D beta 1 and J beta 2.3 loci should have disappeared from this chromosome. The C beta 1 gene indeed was absent from the rearranged chromosome 7, but it was found on chromosome 6 as an inserted segment. The implications of the observations are discussed.
We have isolated a human genomic DNA (designated human frt) cross-hybridizing with the v-ros oncogene of UR2 sarcoma virus. Sequencing analysis of this fragment revealed that this sequence contains a 123-base-pair exon-like structure surrounded by consensus sequences of splice acceptor and donor sites. The deduced amino acid sequence of this stretch was highly homologous to a portion of a tyrosine kinase domain of src family. The human frt sequence was found to be conserved in a wide variety of vertebrates. By using a human-mouse hybridoma panel, human frt was located on chromosome 13, while human c-ros-1 was located on chromosome 6.
We have regionally localized the two human tyrosine kinase proto-oncogenes, c-ros-1 and flt, by in situ hybridization. The c-ros-1 has been mapped to 6q22 and the flt to 13q12.
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An src/yes-related novel gene named syn (SYN in human gene nomenclature) has been identified in the human genome on chromosome 6 and characterized by molecular cloning. Nucleotide sequence analysis of cDNA clones showed that the c-syn gene could encode a protein-tyrosine kinase that is very similar in primary structure to the v-yes and human c-src proteins. A 2.8-kilobase transcript of the c-syn gene, which differs in size from those of the c-yes, c-src, and c-fgr genes, was observed in various cell types. These results show that syn is a new member of the tyrosine kinase oncogene family.
The nucleotide sequence of seven exons of the human c-fgr gene, a cellular homolog of the oncogene of Gardner-Rasheed feline sarcoma virus, was determined. Twenty-six independent genomic clones were obtained from a human gene library with a DNA clone of Y73 avian sarcoma virus oncogene, v-yes, as a probe under relaxed hybridization conditions. Restriction mapping and partial sequence analyses revealed that two of these clones were derived from the c-fgr gene, distinct from the c-yes gene. Interestingly, the splicing points of the c-fgr gene were identical with those of the c-src gene throughout the seven exons, suggesting that the two proto-oncogenes were generated by gene duplication of an ancestral gene containing intervening sequences. On RNA blot hybridization the major transcript was found to be 2.6 kilobase long. Two additional transcripts of 3.5 and 4.7 kilobases were also detected. Furthermore, karyotype analysis of several human-mouse hybrid cells and Southern blot analyses of DNAs of the hybrids with a human c-fgr locus-specific probe showed that this gene is located on chromosome 1.