Mouse Polyoma Virus and Adenovirus Replication in Mouse Cells Temperature-Sensitive in DNA Synthesis

Sheinin, Rose; Fabbro, Julie; Dubsky, Margaret

doi:10.1159/000149638

Cited by 5 publications

(3 citation statements)

References 24 publications

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“…Various experiments have indicated that most known DNA synthesis enzymes including DNA polymerases a, ,B and y, polydeoxynucleotide ligases I and II, poly(ADP-ribose) polymerase and DNA topoisomerase I (reviewed in Sheinin et al, 1983) and DNA topoisomerase H (E.Zacksenhaus, J.Sigouin and R. Sheinin, unpublished results) are unaffected in heat-inactivated ts A1S9 cells. This is in accord with the finding that polyoma viral DNA replication, which is heavily dependent on the host DNA synthesis machinery, is not restricted in temperature-inactivated ts AlS9 cells (Sheinin, 1976b;Sheinin et al, 1985).…”

Section: Introductionsupporting

confidence: 90%

Molecular cloning, primary structure and expression of the human X linked A1S9 gene cDNA which complements the ts A1S9 mouse L cell defect in DNA replication.

Zacksenhaus

Sheinin

1990

The EMBO Journal

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The temperature‐sensitive ts A1S9 mutation of mouse L cells was previously shown to affect nuclear DNA replication and to be complemented by active and inactive human X chromosomes in human‐ts A1S9 somatic cell hybrids. We report the isolation of cDNA clones which correct the ts A1S9 lesion, using as a probe a genomic fragment derived from the human A1S9 locus. The nucleotide sequence of the A1S9 cDNA encompasses a single open reading frame of 2409 bp which could encode a heretofore unreported protein of 90 393 daltons. Southern blot hybridization of the A1S9 cDNA probe with DNA from various species revealed homologous sequences in vertebrates but not in yeast. Northern blot analysis of serum‐starved, synchronized cells demonstrated that the A1S9 gene was expressed at a relatively low level in quiescent cells and at a higher and constant level throughout the cell cycle. Human cell lines harbouring increasing numbers of inactive X chromosomes (47, XXX, 49, XXXXX) were found to express the A1S9 gene at the same level as control cells (45, X), suggesting that the gene does not escape X chromosome inactivation.

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Section: Introductionsupporting

confidence: 90%

Molecular cloning, primary structure and expression of the human X linked A1S9 gene cDNA which complements the ts A1S9 mouse L cell defect in DNA replication.

Zacksenhaus

Sheinin

1990

The EMBO Journal

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“…On the other hand, DNA repair replication proceeds handily even after the ts 2 protein has undergone inactivation. Denaturation of the ts 2 protein does not directly prevent replication of the mouse adenovirus DNA (Sheinin et al, 1985a) or of the linear, double-stranded DNA form of the MoMuLV genome (Richter et al, 1984). On the other hand, synthesis of Py DNA and conversion of the first MoMuLV DNA product to its covalently closed, superhelical intermediate are dependent upon the functional integrity of the ts 2 protein.…”

Section: Discussionmentioning

confidence: 97%

“…Temperature-inactivated ts 2 cells are unable to support replication of polyoma (Py) virus DNA (Slater & Ozer, 1976;Sheinin et al, 1985a). In contrast, they do carry out synthesis of mouse adenovirus DNA (Sheinin et al, 1985a) and of linear, double-stranded Moloney murine leukemia virus (MoMuLV) DNA (Richter et al, 1984).…”

mentioning

confidence: 99%

DNA synthesis in Balb/C-3T3 ts 2 cells is restricted by a temperature-sensitive function of late G1 phase

Sheinin¹,

Mirjah²,

Dubsky³

et al. 1986

Biochemistry

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ts 2 BalB/C-3T3 mouse fibroblasts are cdc mutants, which arrest late in G1, at or near the G1/S traverse, upon full expression of the heat-sensitive lesion. The kinetics of temperature inhibition of DNA synthesis in logarithmically growing cultures reveal three stages of heat inactivation. During the first generation time equivalent, normal semiconservative, semidiscontinuous replication proceeds but is reduced as cells exit and do not reenter S phase. During a second such period, a minimal rate of normal DNA synthesis is maintained. Thereafter, as the cells move into a third aborted cell division cycle, the rate of DNA synthesis increases. However, all semiconservative synthesis is then replaced by DNA repair replication. Temperature inactivation of the ts 2 protein results in shutdown of nuclear DNA synthesis. In contrast, normal replication of mitochondrial DNA proceeds at control rate throughout the first stage of temperature inactivation. Synthesis of this organellar genome is quantitatively reduced as the cells move into the second phase of heat inhibition. Titration of chromatin-bound DNA with ethidium bromide revealed that wild-type cells exhibit a changing DNA topology as the temperature is raised. Temperature-inactivated ts 2 cells behave as though their DNA has been topologically frozen in the configuration of control cells at or near entry into S phase.

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Molecular cloning of human A1S9 locus: An X-linked gene essential for progression through S phase of the cell cycle

Zacksenhaus

Sheinin

1989

Somat Cell Mol Genet

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The temperature-sensitive (ts) A1S9 mouse L-cell mutant is defective in an X-linked gene essential for the progression of cells through the S phase of the cell duplication cycle. We recently reported the complementation of the ts A1S9 cell defect with total human DNA and the isolation of independent temperature-resistant transformants that retained a common set of human specific Alu-containing fragments. Here we describe the molecular cloning of these human DNA sequences from one of the secondary transformants. ST-1-0. A genomic library prepared from ST-1-0 was screened with a total human DNA probe, and two recombinant bacteriophages carrying overlapping segments were isolated. The cloned region was extended in both directions using a human X-chromosome specific library. In total, a human region spanning 42 kb in length, and containing all the Alu-specific DNA sequences found in ST-1-0, was isolated in five overlapping recombinant phages. The A1S9 gene appeared to be larger than the DNA recovered in individual phage isolates, as was assessed by transfection experiments. A single-copy probe derived from the phage DNA was shown to be conserved in independent primary, secondary, and tertiary transformants of ts A1S9 cells and mapped to the X chromosome by molecular hybridization. Northern blot hybridization of this probe with poly(A)+ mRNA derived from ST-1-0 cells identified a transcript of about 3.6 kb.

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Mouse Polyoma Virus and Adenovirus Replication in Mouse Cells Temperature-Sensitive in DNA Synthesis

Cited by 5 publications

References 24 publications

Molecular cloning, primary structure and expression of the human X linked A1S9 gene cDNA which complements the ts A1S9 mouse L cell defect in DNA replication.

Molecular cloning, primary structure and expression of the human X linked A1S9 gene cDNA which complements the ts A1S9 mouse L cell defect in DNA replication.

DNA synthesis in Balb/C-3T3 ts 2 cells is restricted by a temperature-sensitive function of late G1 phase

Molecular cloning of human A1S9 locus: An X-linked gene essential for progression through S phase of the cell cycle

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