Chromosomal assignment of a family of human oncogenes.

A family of human transforming genes, previously shown to share homology with the ras family of viral oncogenes, maps to three different human chromosomes. A well-characterized mouse-human hybrid cell panel, combined with Southern blotting, was used in this study. The transforming gene of the T24 bladder carcinoma cell line maps to human chromosome 11.

Somatic cell genetics and gene families.

The utility of somatic cell genetic analysis for the chromosomal localization of genes in mammals is well established. With the development of recombinant DNA probes and efficient blotting techniques that allow visualization of single-copy cellular genes, somatic cell genetics has been extended from the level of phenotypes expressed by whole cells to the level of the cellular genome itself. This extension has proved invaluable for the analysis of genes not readily expressed in somatic cell hybrids and for the study of multigene families, especially pseudogenes dispersed in different chromosomes throughout the genome.

Expression and stabilization of microinjected plasmids containing the herpes simplex virus thymidine kinase gene and polyoma virus DNA in mouse cells.

To observe the effects of polyoma virus DNA on the expression of the herpes simplex virus (HSV) thymidine kinase (TK) gene early after transfer into TK-deficient mouse cells and the subsequent development of stable TK-positive transformants, we constructed a series of recombinant plasmids containing the herpes simplex virus TK gene joined with various segments of the polyoma virus genome and microinjected them into the nuclei or cytoplasm of LTK-A cells (TK(-), APRT(-)). The frequency of nucleus-injected cells expressing TK after 1 day, measured by autoradiography of cells incubated with [(3)H]thymidine, increased approximately 30-fold when the plasmids contained the polyoma virus origin of replication. The origin includes sequences with homology to the simian virus 40 origin of replication and adjoining sequences, including a recently defined transcription-enhancing sequence.

Genetic control of drug resistance: assignment of ama-1 to Chinese hamster chromosome 7, confirmation of assignment of genes coding for TK, GALK, and ACP to chromosome 7, and tentative assignment of TPI to chromosome 8.

The gene which specifies a subunit of RNA polymerase II, ama-1, is assigned to chromosome 7 in the Chinese hamster. The assignment of genes coding for TK, GALK, and ACP to chromosome 7 is confirmed, with a provisional regional assignment of TK and GALK to 7q. On the basis of one clone with six subclones, a provisional assignment of TPI to Chinese hamster chromosome 8 is made.

Aberrant rearrangement of the kappa light-chain locus involving the heavy-chain locus and chromosome 15 in a mouse plasmacytoma.

The creation of a functional antibody gene requires the precise recombination of gene segments initially separated on the chromosome. Frequently errors occur in the process, resulting in the formation of a non-functional gene. The non-functional genes can be generated by incomplete rearrangements, frameshifts, or the use of pseudo V or J joining segments.

Assignment of the native Chinese hamster dihydrofolate reductase gene to chromosome 2.

The native Chinese hamster DHFR gene was localized to 2pter----q14 using a combination of somatic cell hybrid technology and molecular techniques. In addition, PGD, ENO1, and DTS, previously mapped to Chinese hamster chromosome 2, were localized to 2q14----qter. Localization of PGM1 to this region was confirmed.

The genes coding for the cardiac muscle actin, the skeletal muscle actin and the cytoplasmic beta-actin are located on three different mouse chromosomes.

The actins are a group of highly conserved proteins encoded by a multigene family. We have previously reported that the skeletal muscle actin gene is located on mouse chromosome 3, together with several other unidentified actin DNA sequences. We show here that the gene coding for the cardiac muscle actin, which is closely related to the skeletal muscle actin (1.

Chromosomal assignment of the endogenous proto-oncogene C-abl.

Abelson murine leukemia virus (A-MuLV) is a replication-defective retrovirus that transforms lymphocytes of the B-cell lineage. This virus is a recombinant between the parental Moloney murine leukemia virus and a cellular gene termed C-abl. By analysis of a series of mouse x Chinese hamster hybrid celllines containing various mouse chromosomes, we have mapped the C-abl gene to mouse chromosome 2.

Chromosomal localization of the Moloney sarcoma virus mouse cellular (c-mos) sequence.

The Moloney sarcoma virus-specific onc gene, referred to as v-mos, was used as probe to hybridize to restricted DNAs from various mouse-Chinese hamster hybrid cell lines. These hybrid cells contain, in addition to all of the Chinese hamster chromosomes, various numbers (less than a full complement) of mouse chromosomes. Comparison of the presence or absence of the mouse cellular mos gene with the known karyotype in each of the hybrid cell lines allows us to conclude that the mos gene is on mouse chromosome 4.

DNA sequence associated with chromosome translocations in mouse plasmacytomas.

A DNA sequence that generates aberrantly rearranged immunoglobulin heavy chain constant region genes in murine plasmacytomas is shown to participate in a chromosome translocation. We have previously termed this DNA sequence NIARD for non-immunoglobulin-associated rearranging DNA. NIARD rearrangements were found frequently in murine plasmacytomas but were not detected in normal lymphocytes.

Genetic and biochemical characterization of a human surface determinant on somatic cell hybrids: the 4F2 antigen.

We have mapped the gene which codes the species-specific determinant defined by monoclonal antibody 4F2 to human chromosome 11. All human chromosomes, except Y, were included in a group of four human-mouse hybrid lines. Hybrids heterogeneous for 4F2 antigen expression were sorted using the fluorescence-activated cell sorter (FACS) to yield populations homogeneous with respect to the presence or absence of this determinant.

Identification of human genomic clones coding the major histocompatibility antigens HLA-a2 and HLA-B7 by DNA-mediated gene transfer.

We have screened a large number of isolated human genomic clones that hybridize to a cloned HLA cDNA probe for their ability to direct the synthesis of HLA-A, -B, and -C surface antigens on mouse L cells following DNA-mediated gene transfer. The surface expression of human histocompatibility antigens, monitored by indirect immunofluorescence and the fluorescence-activated cell sorter, was examined at 60 hr after transfection and on hypoxanthine/aminopterin/thymidine-resistant (HATR) populations derived from cotransfer with the herpes simplex virus thymidine kinase gene. Two unique genomic clones designated JY B3.

Multiple human beta interferon genes.

Analysis of human beta interferon (IFN) mRNA preparations obtained from poly(I) . poly (C)-induced human diploid fibroblasts (FS-4) and from several similarly induced human-mouse somatic cell hybrids by electrophoresis through agarose-CH3HgOH tube gels led to the detection of at least five translationally active human IFN-beta mRNA species. The results obtained are consistent with the existence of IFN-beta genes on different human chromosomes.

Structural genes of the mouse major urinary protein are on chromosome 4.

The major urinary proteins (MUPs) of mouse are a family of at least three major proteins which are synthesized in the liver of all strains of mice. The relative levels of synthesis of these proteins with respect to each other in the presence of testosterone is regulated by the Mup-a locus located on chromosome 4. In an effort to determine the mechanism of this regulation in molecular terms, a cDNA clone containing most of the coding region of a MUP protein has been isolated and identified by partial DNA sequence analysis.

Dispersion of argininosuccinate-synthetase-like human genes to multiple autosomes and the X chromosome.

DNA sequences closely homologous to argininosuccinate synthetase are present at ten or more distinct locations in the human genome, including sites on chromosomes 6, 9 and X. Argininosuccinate synthetase thus represents one of the most widely dispersed multigene families described to date, the first instance of a multigene family associated with an enzyme of intermediary metabolism and, perhaps most striking, the first instance of a multigene family with members on both autosomes and sex chromosomes.

Partial purification and characterization of the mRNA for human thymidine kinase and hypoxanthine/guanine phosphoribosyltransferase.

We used direct microinjection of poly(A)+RNA into individual hypoxanthine/guanine phosphoribosyltransferase-deficient or thymidine kinase-deficient cells and detected the specific in vivo translation products as an assay for human hypoxanthine/guanine phosphoribosyltransferase or thymidine kinase mRNAs. The incorporation of [3H]hypoxanthine or [3H]thymidine into cells in response to injected mRNA was assayed in situ by autoradiography. Methylmercuric hydroxide/agarose gel analysis showed that human hypoxanthine/guanine phosphoribosyltransferase mRNA contains approximately 1,530 nucleotides, which is twice the number required for its protein coding capacity.

Somatic cell genetic analysis of HLA-A, B, C and human beta 2-microglobulin expression.

We have examined the cell surface expression of the human histocompatibility antigens HLA-A, B, C and beta 2-microglobulin (beta 2m) on a human-mouse somatic cell hybrid line. Using specific antibodies and the fluorescence-activated cell sorter (FACS), we viably fractionated and characterized four separate hybrid subpopulations (HLA+,beta 2m+; HLA+,beta 2m-; HLA-,beta 2m+; HLA-,beta 2m-). Hybrid selection based on surface antigen expression resulted in corresponding genetic selection for and against human chromosomes 6 and 15.

Human beta interferon gene localization and expression in somatic cell hybrids.

The human fibroblast interferon gene beta 1 was mapped to human chromosome 9. Sequence homology with a beta 1 cDNA clone was detected in both genomic DNA and induced mRNA of human/mouse or human/hamster somatic cell hybrids containing human chromosome 9, but not in lines lacking this chromosome or those retaining a complex translocation involving chromosomes 9 and 11. Interferon mRNA that did not share sequence homology with the beta 1 cDNA clone was detected in lines containing human chromosomes 2 and 5 but lacking chromosome 9, suggesting the presence of other unlinked interferon sequences in the human genome.

Localization of the casein gene family to a single mouse chromosome.

A series of mouse-hamster somatic cell hybrids containing a variable number of mouse chromosomes and a constant set of hamster chromosomes have been used to determine the chromosomal location of a family of hormone-inducible genes, the murine caseins. Recombinant mouse cDNA clones encoding the alpha-, beta-, and gamma-caseins were constructed and used in DNA restriction mapping experiments. All three casein cDNAs hybridized to the same set of somatic cell hybrid DNAs isolated from cells containing mouse chromosome 5, while negative hybridization was observed to ten other hybrid DNAs isolated from cells lacking chromosome 5.

Chromosomal location of a human alpha interferon gene family.

To determine the chromosomal location of the human alpha interferon genes, we scored a series of human/rodent somatic cell hybrids for the presence of DNA sequences hybridizing to an alpha 1 interferon DNA probe. The presence of human chromosome 9 in a hybrid correlated with the presence of a family of alpha interferon genes.

J chain is encoded by a single gene unlinked to other immunoglobulin structural genes.

Immunoglobulin J chain mediates the polymerization of both IgM and IgA immunoglobulins. Its synthesis is closely regulated in B lymphocytes, apparently at the level of RNA transcription. To define the genetic bases of this regulation, we have determined the location and number of J chain genes in the mouse.