Effects of 5-bromodeoxyuridine on cell division and DNA replication in HeLa cells*1, *2, *3

Simon, Edward H.

doi:10.1016/0014-4827(63)90268-4

Cited by 43 publications

(12 citation statements)

References 4 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The use of FUdR raises the possibility that the quantity or quality of BUdR incorporation into DNA may be different from that obtained when endogenous thymidylate synthesis is not prevented . BUdR incorporation has been reported to be enhanced in the presence of FUdR (Dvordjevic and Szybalski, 1960), although Simon (1963) and Littlefield and Gould (1960) did not observe this effect . Also, thymidylate deprivation may produce DNA lesions that would be susceptible to nonsemiconservative BUdR incorporation via a repair mechansim (Pauling and Hanawalt, 1965) .…”

Section: Discussionmentioning

confidence: 96%

“…It has not been determined, however, whether incorporation of the analogue during one round of DNA synthesis is sufficient to block fusion of myoblasts during the following G1 . Since replication in BUdR is accompanied by semiconservative distribution of the newly synthesized DNA (Simon, 1963 ;Haut and Taylor, 1967), growth in the analogue for one S period should produce daughter cells that have one normal and one BU-containing polynucleotide strand in the DNA duplex .…”

Section: Effect Of Unifilar Incorporation Of Bummentioning

confidence: 99%

See 1 more Smart Citation

Inhibition of Myoblast Fusion After One Round of Dna Synthesis in 5-Bromodeoxyuridine

Bischoff

Holtzer

1970

The Journal of Cell Biology

311

View full text Add to dashboard Cite

The thymidine analogue 5-bromodeoxyuridine (BUdR) has a differential effect on the synthesis of tissue-specific products and molecules required for growth and division . Proliferating myogenic cells cultured in BUdR fail to fuse and fail to initiate the synthesis of contractile protein filaments . Conversely, BUdR has but a minor effect on cell viability and reproductive integrity . Low concentrations of BUdR result in an enhancement of cell number relative to the controls ; higher concentrations are cytotoxic . Suppression of myogenesis is reversible after at least 10 cell generations of growth in the analogue . Cells that do not synthesize DNA, such as postmitotic myoblasts and myotubes, are not affected by BUdR . Incorporation of BUdR for one round of DNA synthesis was accomplished by first incubating myogenic cells, prior to fusion, in 5-fluorodeoxyuridine (FUdR) to block DNA synthesis and collect cells in the presynthetic phase . The cells were then allowed to synthesize either normal DNA or BU-DNA for one S period by circumventing the FUdR block with BUdR or BUdR plus thymidine (TdR) . The cultures were continued in FUdR to prevent dilution of the incorporated analogue by further division . After 3 days, the cultures from the FUdR-BUdR series showed the typical BUdR effect ; the cells were excessively flattened and few multinucleated myotubes formed . Cells in the control cultures were of normal morphology, and multinucleated myotubes were present . These results were confirmed in another experiment in which BUdR3H was added to 2-day cultures in which myotubes were forming. Fusion of thymidine 3H-labeled cells begins at 8 hr after the preceding S phase . In contrast, cells which incorporate BUdR-3H for one S period do not fuse with normal myotubes.

show abstract

Section: Discussionmentioning

confidence: 96%

Section: Effect Of Unifilar Incorporation Of Bummentioning

confidence: 99%

Inhibition of Myoblast Fusion After One Round of Dna Synthesis in 5-Bromodeoxyuridine

Bischoff

Holtzer

1970

The Journal of Cell Biology

311

View full text Add to dashboard Cite

show abstract

“…Chinese hamster cells in these experiments, however, showed no selectivity for TdR or BrUdR during either semiconservative or repair replication. Once incorporated into DNA, however, there is ample evidence which shows that BrUdR inhibits cell growth [24,25] and produces many abnormalities . But in the initial incorporation into DNA, there does not seem to be any obvious discrimination between TdR and BrUdR.…”

Section: Discussionmentioning

confidence: 99%

Repair Replication in Chinese Hamster Cells After Damage From Ultraviolet Light*

Cleaver

1970

Photochem & Photobiology

View full text Add to dashboard Cite

After irradiation with U.V. light, Chinese hamster cells perform repair replication of their DNA. Small numbers of nucleosides are inserted into DNA, such that when BrUdR is used there is no detectable change in density. Repair replication begins immediately after irradiation, but it decelerates steadily and at least half is complete within 4 hr. Repair replication saturates above 200ergs/mmz at a level which represents 0.055 per cent replacement of all thymine sites in 4 hr. Repair replication in mammalian cells, in contrast to that in microorganisms, does not appear to replace pyrimidine dimers excised from DNA in acid soluble form, and neither repair nor semiconservative replication discriminates between BrUdR and TdR.

show abstract

“…To define whether the block in productive proliferation due to loss of CAML occurs prior to this S phase transition, we conducted a cell cycle assay in vitro by labeling cells with BrdU and 7-aminoactinomycin D for 30 min after a 72-h period of TCR stimulation (27,28). Cell cycle analysis showed that tCAML 2/2 T cells incorporated BrdU highly efficiently, and increased their DNA content, indicating that they were able to enter the S and G 2 phases of the cell cycle (Fig.…”

Section: Caml-deleted T Cells Enter the S Phase Of The Cell Cyclementioning

confidence: 99%

Calcium-Modulating Cyclophilin Ligand Is Essential for the Survival of Activated T Cells and for Adaptive Immunity

Chan

Lindquist

Hansen

et al. 2015

The Journal of Immunology

View full text Add to dashboard Cite

Calcium-modulating cyclophilin ligand (CAML) is an endoplasmic reticulum resident protein that is widely expressed. Although it has been demonstrated to participate in the tail-anchored protein insertion pathway, its physiological role in the mature immune system is unknown. In this work, we show that mature, peripheral T cells require CAML for survival specifically following TCR-induced activation. In this study, we examined mature T cells from spleen and lymph nodes of tamoxifen-inducible CAML knockout mice (tCAML−/−). Whereas CAML-deficient T cells were able to express the early activation markers CD25 and CD69, and produce IL-2 normally upon stimulation, deficient cells proliferated less and died. Cells did not require CAML for entry into the S phase of the cell cycle, thus implicating its survival function at a relatively late step in the T cell activation sequence. In addition, CAML was required for homeostatic proliferation and for Ag-dependent cell killing in vivo. These results demonstrate that CAML critically supports T cell survival and cell division downstream of T cell activation.

show abstract

Effects of 5-bromodeoxyuridine on cell division and DNA replication in HeLa cells1, 2, *3

Cited by 43 publications

References 4 publications

Inhibition of Myoblast Fusion After One Round of Dna Synthesis in 5-Bromodeoxyuridine

Inhibition of Myoblast Fusion After One Round of Dna Synthesis in 5-Bromodeoxyuridine

Repair Replication in Chinese Hamster Cells After Damage From Ultraviolet Light*

Calcium-Modulating Cyclophilin Ligand Is Essential for the Survival of Activated T Cells and for Adaptive Immunity

Contact Info

Product

Resources

About