Effects of aphidicolin and hydroxyurea on the cell cycle and differentiation of Trypanosoma brucei bloodstream forms

Mutomba, Martha; Wang, Ching C.

doi:10.1016/0166-6851(96)02675-8

Cited by 42 publications

(25 citation statements)

References 31 publications

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“…The conclusion is in accordance with the sensitivity of trypanosomes towards hydroxyurea [17][18][19] although at low concentrations of hydroxyurea inhibition of ribonucleotide reductase seems not to be the primary effect [19]. The sensitivity of ribonucleotide reductases from different species towards radical scavengers varies remarkably and may be due to the accessibility of the iron/radical center [14].…”

Section: Sequence Comparison Of 72 Brucei R2 With Other R2 Proteinssupporting

confidence: 65%

Cloning, sequencing and expression of ribonucleotide reductase R2 from Trypanosoma brucei

et al. 1997

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As a first step towards the understanding of the trypanosomal synthesis of deoxyribonucleotides we describe the cloning of the gene of ribonucleotide reductase subunit R2 from 72 brucei hrucei and its overexpression in E. coli. Materials and methods DNA and RNA preparation2;4 l08 long slender 72 brucei cells were suspended in 10 mM Tris-HC1, 250 mM NaCI, 0.5% Nonidet P-40, pH 8.0. After 5 rain incubation on ice the suspension was centrifuged and the cell pellet was homogenized in 200 lal 10 mM Tris-HC1, 10 mM NaC1, 10 mM EDTA, 0.5% SDS, pH 8.0. 50 lag proteinase K were added and the mixture was incubated for 1 h at 37°C. DNA was purified by phenol extraction.RNA was isolated from bloodstream long slender and short stumpy as well as procyclic stages of 72 brucei using the RNA Easy Kit (Qiagen) according to the instructions of the manufacturer.

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Section: Sequence Comparison Of 72 Brucei R2 With Other R2 Proteinssupporting

confidence: 65%

Cloning, sequencing and expression of ribonucleotide reductase R2 from Trypanosoma brucei

et al. 1997

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“…The SVK14 and G361 cells were prepared as described above by seeding cells into 25 cm 2 Acyclovir (Cioe et al, 1992 (Mutomba and Wang, 1996) Early S 0.1 µg ml (Doyle et al, 1996) G 0 /G 1 4°C 0 0 Mimosine (Hoffman et al, 1991) G 1 300 µM (Morice et al, 1993) G 1 0.1 nm D 0 0 Staurosporine (Zong et al, 1999) G 1 20 nm…”

Section: Screening Of Cell Cycle Synchronizing Agentsmentioning

confidence: 99%

Effect of sulphur mustard on human skin cell lines with differential agent sensitivity

Simpson

Lindsay

2005

J of Applied Toxicology

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The ability of sulphur mustard (HD) to induce DNA damage places limits on the efficacy of approaches aimed at protecting human cells from the cytotoxic effects of HD using a variety of protective agents such as thiol-containing esters and protease inhibitors. In the present study, potential alternative strategies were investigated by examining the differential effects of HD on G361, SVK14, HaCaT and NCTC 2544 human skin cells. The G361 cell line was more resistant to the cytotoxic effects of HD than the NCTC, HaCaT and SVK14 cell lines at HD doses of >3 and <100 microM HD as determined by the MTT assay. At 72 h after exposure to 60 microM HD there was up to an 8.8-fold difference (P < 0.0001) between G361 and SVK14 cell culture viability. Buthionine sulphoximine (BSO) pretreatment increased the sensitivity of all four cell lines to HD. A substantial proportion of the resistance of G361 cells to HD was attributable to BSO-mediated effects on antioxidant-mediated metabolism, although G361 cultures still retained a high degree of viability at 30 microM HD following BSO pretreatment. Cell cycle analysis confirmed that SVK14 cells were relatively more sensitive to HD, as shown by the 2.1-fold reduction (P < 0.0001) in the percentage of cells in G0/G1 phase 24 h after HD exposure compared with control cultures. This compared well with a 1.2-fold increase (P < 0.05) in the percentage of G361 cells in G0/G1 phase following HD exposure, suggesting the existence of a more efficient G0/G1 checkpoint control mechanism in this cell line. Manipulation of the cell cycle using various modulating agents did not increase the resistance of cell lines to the cytotoxic effects of HD.

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“…L'effet de HU est réversible après lavage des cellules et remise en culture dans du milieu neuf. Les cellules entrent alors de façon synchronisée en phase S (Mutomba et Wang, 1996). Notons, toutefois, que les effets de HU peuvent être très différents en fonction des cellules et de la concentration utilisée.…”

Section: ) Répartition Des Cellules Shep Dans Le Cycle Cellulaireunclassified

“…Bien qu'ils utilisent des lignées de la même famille de cellules, des neuroblastomes, Fulda et al (1995) notent des résultats assez variables sur l'action des agents cytobloquants d'une lignée à l'autre. De même, Mutomba et Wang (1996), montrent que pour certains types de cellules HU n'inhibe pas la synthèse d'ADN. Elle les bloque plutôt à la transition G2+M ou M/G1.…”

Section: ) Répartition Des Cellules Shep Dans Le Cycle Cellulaireunclassified

Internal dynamics of a living cell nucleus investigated by dynamic light scattering

et al. 2008

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Effects of aphidicolin and hydroxyurea on the cell cycle and differentiation of Trypanosoma brucei bloodstream forms

Cited by 42 publications

References 31 publications

Cloning, sequencing and expression of ribonucleotide reductase R2 from Trypanosoma brucei

Cloning, sequencing and expression of ribonucleotide reductase R2 from Trypanosoma brucei

Effect of sulphur mustard on human skin cell lines with differential agent sensitivity

Internal dynamics of a living cell nucleus investigated by dynamic light scattering

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