Isolation of temperature sensitive mammalian cells by selective detachment

Roscoe, D.H.; Read, Moira; Robinson, Hildred

doi:10.1002/jcp.1040820302

Cited by 53 publications

(18 citation statements)

References 15 publications

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“…The Chinese hamster fibroblast cell lines WglA and K12 have been described previously (9,17). K12 GRP94, respectively, has been described previously (11,12).…”

Section: Methodsmentioning

confidence: 99%

Calcium ionophore A23187 induces expression of glucose-regulated genes and their heterologous fusion genes.

Resendez¹,

Attenello²,

Grafsky³

et al. 1985

Mol. Cell. Biol.

148

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Using two cDNA clones which encode hamster genes specifically induced by glucose starvation, we demonstrated that an 8- and 30-fold increase, respectively, in the transcription rates of these genes was coordinately effected by calcium ionophore A23187 treatment, resulting in a similar increase in the steady-state levels of their mRNAs. This response was observed within several hours of ionophore treatment in several mammalian cell types and appeared to be specifically mediated by A23187 but not by other ionophores in general. To define the regulatory sequence which mediates this Ca2+-induced response, we showed by gene transfection techniques that the 5' flanking sequence of a rat glucose-regulated gene contained the region for induction by A23187. The system reported here offers attractive features for the study of specific gene regulation by Ca2+.

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“…The Chinese hamster fibroblast cell lines WglA and K12 have been described previously (9,17). K12 GRP94, respectively, has been described previously (11,12).…”

Section: Methodsmentioning

confidence: 99%

Calcium ionophore A23187 induces expression of glucose-regulated genes and their heterologous fusion genes.

Resendez¹,

Attenello²,

Grafsky³

et al. 1985

Mol. Cell. Biol.

148

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“…First, the transformation state of the cells may influence the cell cycle-dependent activity of transcription factors. The K12 cells used in this study were derived from an established Chinese hamster lung fibroblast cell line (25), while the AEV ts34 cells used by Dalton A second explanation is that cells were synchronized by different methods in the two studies. Dalton and Wells blocked cells in the beginning of S phase with aphidicolin, a reversible inhibitor of DNA polymerase a; we used serum starvation to block cells in early G1 phase.…”

Section: [Ccaat Mutation]mentioning

confidence: 99%

Cell cycle regulation of H2b histone octamer DNA-binding activity in Chinese hamster lung fibroblasts.

Ito¹,

Sharma²,

Lee³

et al. 1989

Mol. Cell. Biol.

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The promoter regions of H2b histone genes contain a 14-base-pair element which includes the octamer ATTTGCAT. Mutational analysis has implicated the octamer element in the cell cycle-dependent expression of H2b histone genes. In this report, we address the question of whether the DNA-binding activity of the octamer transcription factor is itself cell cycle regulated. By using a gel mobility shift assay, we measured the relative amounts of octamer-binding activity during various phases of the cell cycle in serum-synchronized Chinese hamster fibroblasts. We found that the activity increased approximately fivefold between late Gl phase and early S phase and then decreased threefold between late S phase and G2 phase. These cell cycle-dependent changes in octamer DNA-binding activity may in part account for the selective transcription of H2b histone genes in late Gl and S phases.

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“…In mammalian cells (Roscoe et al ., 1973;Talavera and Basilico, 1977), bacteria (Roscoe et al ., 1973), viruses (Fields and Joklik, 1969;Roscoe et al ., 1973) and yeast (Hartwell et al ., 1970;Hartwell, 1974), loss-of-function mutants have been utilized to explore the regulation of events within the eukaryotic cell cycle. In T. gondii , tachyzoite temperature-sensitive mutants have been generated using a chemical mutagenesis strategy (Pfefferkorn and Pferfferkorn, 1976;Radke et al ., 2000).…”

Section: Introductionmentioning

confidence: 99%

Genetic rescue of a Toxoplasma gondii conditional cell cycle mutant

White

Jerome

Vaishnava

et al. 2005

Molecular Microbiology

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SummaryGrowth rate is a major pathogenesis factor in the parasite Toxoplasma gondii ; however, how cell division is controlled in this protozoan is poorly understood. Herein, we show that centrosomal duplication is an indicator of S phase entry while centrosome migration marks mitotic entry. Using the pattern of centrosomal replication, we confirmed that mutant ts 11C9 undergoes a bimodal cell cycle arrest that is characterized by two subpopulations containing either single or duplicated centrosomes which correlate with the bipartite genome distribution observed at the non-permissive temperature. Genetic rescue of ts 11C9 was performed using a parental RH strain cDNA library, and the cDNA responsible for conferring temperature resistance (growth at 40 ∞ ∞ ∞ ∞ C) was recovered by recombination cloning. A single T. gondii gene encoding the protein homologue of XPMC2 was responsible for genetic rescue of the temperature-sensitive defect in ts 11C9 parasites. This protein is a known suppressor of mitotic defects, and in tachyzoites, TgXPMC2-YFP localized to the parasite nucleus and nucleolus which is consistent with the expected subcellular localization of critical mitotic factors. Altogether, these results demonstrate that ts 11C9 is a conditional mitotic mutant containing a single defect which influences two distinct control points in the T. gondii tachyzoite cell cycle.

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Isolation of temperature sensitive mammalian cells by selective detachment

Cited by 53 publications

References 15 publications

Calcium ionophore A23187 induces expression of glucose-regulated genes and their heterologous fusion genes.

Calcium ionophore A23187 induces expression of glucose-regulated genes and their heterologous fusion genes.

Cell cycle regulation of H2b histone octamer DNA-binding activity in Chinese hamster lung fibroblasts.

Genetic rescue of a Toxoplasma gondii conditional cell cycle mutant

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