Species Specificity of an Enzyme Determined by an Erythrocyte Nucleus in an Interspecific Hybrid Cell

1970 ◽  
Vol 7 (1) ◽  
pp. 1-3
Author(s):  
P. R. COOK
Keyword(s):  
Interspecific Hybrid ◽  
Mutant Mouse ◽  
Hybrid Cell ◽  
Mouse Cell ◽  
Species Specificity ◽  
Inosinic Acid ◽  
Erythrocyte Nucleus

When a chick erythrocyte nucleus is introduced into the cytoplasm of a mutant mouse cell lacking inosinic acid pyrophosphorylase, synthesis of the enzyme is induced. The enzyme induced in this way has the characteristics of chick, not mouse, inosinic acid pyrophosphorylase.

SYNTHESIS OF AN ENZYME DETERMINED BY AN ERYTHROCYTE NUCLEUS IN A HYBRID CELL

1969 ◽  
Vol 5 (1) ◽  
pp. 121-133
Author(s):  
H. HARRIS ◽  
P. R. HARRIS
Keyword(s):  
Mutant Mouse ◽  
Hybrid Cell ◽  
Critical Role ◽  
Surface Antigens ◽  
Mouse Cell ◽  
Inosinic Acid ◽  
Erythrocyte Nucleus ◽  
Transfer Of Information ◽  
Kinetics Of

When a chick erythrocyte nucleus is introduced into the cytoplasm of a mutant mouse cell lacking the enzyme inosinic acid pyrophosphorylase, synthesis of the enzyme is induced. However, this synthesis does not begin until the erythrocyte nucleus develops a nucleolus. The kinetics of synthesis of the enzyme are essentially similar to those previously described for the synthesis of surface antigens specified by the chick nucleus. The results of both sets of experiments indicate that the nucleolus plays a critical role in the transfer of information from the genes to the cytoplasm of the cell.

scholarly journals The 27-kD diphtheria toxin receptor-associated protein (DRAP27) from vero cells is the monkey homologue of human CD9 antigen: expression of DRAP27 elevates the number of diphtheria toxin receptors on toxin-sensitive cells.

1992 ◽  
Vol 118 (6) ◽  
pp. 1389-1399 ◽  
Author(s):  
T Mitamura ◽  
R Iwamoto ◽  
T Umata ◽  
T Yomo ◽  
I Urabe ◽  
...  
Keyword(s):  
Diphtheria Toxin ◽  
Binding Capacity ◽  
Hybrid Cell ◽  
Antigen Expression ◽  
Vero Cells ◽  
Mouse Cell ◽  
Homology Search ◽  
Receptor Associated Protein ◽  
New Family ◽  
Cloned Cdna

Diphtheria toxin (DT) receptor associates with a 27-kD membrane protein (DRAP27) in monkey Vero cells. A cDNA encoding DRAP27 was isolated, and its nucleotide sequence was determined. The deduced amino acid sequence revealed that DRAP27 is the monkey homologue of human CD9 antigen. DRAP27 is recognized by CD9 antibodies. A human-mouse hybrid cell line (3279-10) possessing human chromosome 5, sensitive to DT, but not expressing CD9 antigen, was used for transfection experiments with DRAP27. When the cloned cDNA encoding DRAP27 was transiently expressed in 3279-10 cells, the total DT binding capacity was three to four times higher than that of untransfected controls. Transfectants stably expressing DRAP27 have an increased number of DT binding sites on the cell surface. Furthermore, the transfectants are 3-25 times more sensitive to DT than untransfected cells, and the sensitivity of these cells to DT is correlated with the number of DRAP27 molecules on the surface. However, when the cloned cDNA was introduced into mouse cell lines that do not express DT receptors, neither an increased DT binding nor enhancement of DT sensitivity was observed. Hence, we conclude that DRAP27 itself does not bind DT, but serves to increase DT binding and consequently enhances DT sensitivity of cells that have DT receptors. 12 proteins related to DRAP27/CD9 antigen were found through homology search analysis. These proteins appear to belong to a new family of transmembrane proteins.

Correction of Genetic Defects in Mammalian Cells by the Input of Small Amounts of Foreign Genetic Material

1973 ◽  
Vol 13 (3) ◽  
pp. 841-861
Author(s):  
YVONNE L. BOYD ◽  
H. HARRIS
Keyword(s):  
Thymidine Kinase ◽  
Mammalian Cells ◽  
Hybrid Cell ◽  
Genetic Material ◽  
Chinese Hamster ◽  
Heat Sensitivity ◽  
Chinese Hamster Cells ◽  
Inosinic Acid ◽  
Mouse Cells

Chinese hamster cells lacking inosinic acid pyrophosphorylase and mouse cells lacking thymidine kinase were fused with chick erythrocytes. The resultant heterokaryons were cultivated in a selective medium in which possession of these enzymes was essential for cell survival and growth. Clones of cells able to grow in this medium were isolated and studied. A detailed karyological analysis of these clones failed to reveal any chick chromosomes; nor could any chick-specific antigens be detected on the surface of the cells. Nonetheless, clones arising from the fusion of chick erythrocytes with Chinese hamster cells were shown to possess an inosinic acid pyrophosphorylase which had the electrophoretic characteristics of chick inosinic acid pyrophosphorylase. However, the clones arising from the fusion of the chick erythrocytes with the mouse cells had a thymidine kinase with the electrophoretic mobility and heat sensitivity of murine, not chick, thymidine kinase. Both types of hybrid cell have now been cultivated in vitro for 18 months without the loss of thymidine kinase or inosinic acid pyrophosphorylase activity.

The Expression of Genetic Information: A Study with Hybrid Animal Cells

1969 ◽  
Vol 4 (2) ◽  
pp. 499-525
Author(s):  
H. HARRIS ◽  
E. SIDEBOTTOM ◽  
D. M. GRACE ◽  
M. E. BRAMWELL
Keyword(s):  
Specific Surface ◽  
Genetic Information ◽  
Decisive Role ◽  
Surface Antigens ◽  
Mouse Cell ◽  
Animal Cells ◽  
Hybrid Animal ◽  
Specific Antigens ◽  
Erythrocyte Nucleus ◽  
Transfer Of Information

When the nucleus of a hen erythrocyte is introduced into the cytoplasm of a human or mouse cell in culture, it resumes the synthesis of RNA. The reactivated erythrocyte nucleus undergoes great enlargement, but it does not, for at least 2 or 3 days, develop nucleoli which can be discerned under the light microscope. During this period, the heterokaryon, although it may contain several active erythrocyte nuclei, does not synthesize any hen-specific surface antigens; and the hen-specific antigens introduced into the surface of the heterokaryon by the process of cell fusion are eliminated. But when, later, the erythrocyte nuclei do develop nucleoli, hen-specific antigens reappear on the surface of the heterokaryon and progressively accumulate. Before developing nucleoli, the erythrocyte nuclei synthesize little, if any, normal 28 S or 16 S RNA; but they do synthesize large amounts of the RNA which shows polydisperse sedimentation in conventional sucrose density gradients. Autoradiographic studies involving the use of a microbeam of ultraviolet light show, however, that this ‘polydisperse’ RNA is not transferred to the cytoplasm of the cell in detectable amounts so long as the erythrocyte nucleus lacks a definitive nucleolus. The inability of the erythrocyte nucleus at this stage to determine the synthesis of hen-specific surface antigens is thus attributable to the fact that it fails to transfer the RNA made on its chromosomes to the cytoplasm of the cell. When the erythrocyte nuclei develop nucleoli, however, the RNA which they make is transferred to the cytoplasm of the cell, and the synthesis of hen-specific surface antigens then begins. These experiments suggest that the nucleolus may play a decisive role in the transfer of information from nucleus to cytoplasm. The possible nature of this role is discussed.

scholarly journals Identification of the Myelin Protein Plasmolipin as the Cell Entry Receptor for Mus caroli Endogenous Retrovirus

2008 ◽  
Vol 82 (14) ◽  
pp. 6862-6868 ◽  
Author(s):  
A. Dusty Miller ◽  
Ulla Bergholz ◽  
Marion Ziegler ◽  
Carol Stocking
Keyword(s):  
Cell Line ◽  
Hybrid Cell ◽  
Receptor Gene ◽  
Wild Mouse ◽  
Cell Types ◽  
Mouse Cell ◽  
Endogenous Retrovirus ◽  
Receptor Activity ◽  
Endogenous Virus ◽  
Mus Caroli

ABSTRACT The Asian wild mouse species Mus caroli harbors an endogenous retrovirus (McERV) that is closely related to but distinct from the endogenous retrovirus family defined by the Mus dunni endogenous virus and the Mus musculus endogenous retrovirus. McERV could infect some cell types from humans, dogs, and rats, but not all, and did not infect any mouse cell line tested. Because of its interesting host range and proposed ancestral relationship to primate retroviruses and because none of the entry receptors for this family of retroviruses had been identified, we began a search for the McERV receptor. We determined the chromosomal location of the receptor gene in the human genome by phenotypic screening of the G3 human-hamster radiation hybrid cell line panel and confirmed the localization by assaying for receptor activity conferred by bacterial artificial chromosome (BAC) clones spanning the region. We next localized the gene more precisely in one positive BAC by assaying for receptor activity following BAC digestion with several restriction enzymes that cleaved different sets of genes, and we confirmed that the final candidate gene, plasmolipin (PLLP; TM4SF11), is the novel receptor by showing that the expression of the human PLLP cDNA renders hamster and mouse cells susceptible to McERV infection. PLLP functions as a voltage-dependent potassium ion channel and is expressed primarily in kidney and brain, helping to explain the limited range of cell types that McERV can infect. Interestingly, mouse PLLP also functioned well as a receptor for McERV but was simply not expressed in the mouse cell types that we originally tested.

scholarly journals Host-Specific Replication of BK Virus DNA in Mouse Cell Extracts Is Independently Controlled by DNA Polymerase α-Primase and Inhibitory Activities

2010 ◽  
Vol 84 (13) ◽  
pp. 6636-6644 ◽  
Author(s):  
Irina Tikhanovich ◽  
Heinz Peter Nasheuer
Keyword(s):  
Dna Replication ◽  
Dna Polymerase ◽  
Mouse Cell ◽  
Bk Virus ◽  
Species Specificity ◽  
Human Polyomavirus ◽  
Dna Polymerase Α ◽  
The Core ◽  
Inhibitory Activities ◽  
Species Specific

ABSTRACT The activation of the human polyomavirus BK causes polyomavirus-associated nephropathy in immunocompromised humans. Studies of the virus have been restricted since the virus DNA replication is species specific. Cell-based and cell-free DNA replication systems, including the BK virus (BKV) monopolymerase DNA replication system using purified proteins, reproduce the species specificity (28). Therefore, the major host proteins comprising this assay, DNA polymerase α-primase (Pol-prim) and replication protein A (RPA), were intensively studied here. We demonstrate that Pol-prim plays a major role in the species specificity of BKV DNA replication. Both large subunits p180 and p68 of the enzyme complex have central functions in modulating the host specificity. Recently, an inhibitory activity of BKV DNA replication was described (C. Mahon, B. Liang, I. Tikhanovich, J. R. Abend, M. J. Imperiale, H. P. Nasheuer, and W. R. Folk, J. Virol. 83:5708-5717, 2009), but neither mouse Pol-prim nor mouse RPA diminishes cell-free BKV DNA replication. However, the inhibition of BKV DNA replication in mouse extracts depends on sequences flanking the core origin. In the presence of human Pol-prim, the inhibitory effect of mouse cell factors is abolished with plasmid DNAs containing the murine polyomavirus early promoter region, whereas the late enhancer region and the core origin are supplied from BKV. Thus, BKV replication is regulated by both Pol-prim, as a core origin species-specific factor, and inhibitory activities, as origin-flanking sequence-dependent factor(s).

Recovery from post-irradiation inhibition of DNA synthesis in an ultraviolet-sensitive mutant mouse cell

1982 ◽  
Vol 104 (4-5) ◽  
pp. 305-309 ◽  
Author(s):  
Hiroko Hama-Inaba ◽  
Naoko Hieda-Shiomi ◽  
Tadahiro Shiomi ◽  
Koki Sato
Keyword(s):  
Dna Synthesis ◽  
Mutant Mouse ◽  
Mouse Cell ◽  
Sensitive Mutant ◽  
Post Irradiation ◽  
Inhibition Of Dna Synthesis
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