The relation between 5-amino uracil-induced mitotic synchronization and DNA synthesis

Jakob, Karl M.; Trosko, James E.

doi:10.1016/0014-4827(65)90289-2

Cited by 39 publications

(11 citation statements)

References 8 publications

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“…DNA synthesis was measured over time using 5-aminouracil, which synchronizes DNA synthesis during cell division (Jakob and Trosko, 1965). The level of DNA synthesis, as measured by 3 H-thymidine incorporation into synchronized roots, was not significantly different between control and mutant plants.…”

Section: Dna Synthesis In 35s-psugt1 Antisense Plants Is Retarded Commentioning

confidence: 99%

“…Seeds of Pisum sativum cv Little Marvel (Royal Seed Company, Kansas City, MO) were surface-sterilized by immersion in 95% (v/v) eth- Cell division in roots was synchronized by treatment with 12 mM 5-aminouracil for 2 hr (Jakob and Trosko, 1965). DNA synthesis in roots of control plants (dashed line) or plants expressing 35S-PsUGT1 antisense cDNA (solid line) was measured in counts per minute (cpm) by pulse labeling with 3 H-thymidine every 3 hr.…”

Section: Plant Growthmentioning

confidence: 99%

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Meristem-Localized Inducible Expression of a UDP-Glycosyltransferase Gene Is Essential for Growth and Development in Pea and Alfalfa

et al. 1999

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PsUGT1 , which encodes a microsomal UDP-glucuronosyltransferase, was cloned from root tips of Pisum sativum . PsUGT1 expression is correlated with mitosis and strongly induced in dividing cells. A region at the C terminus of the encoded protein is closely related to the UDP-glucuronic acid binding site consensus sequence, and the protein encoded by PsUGT1 catalyzes conjugation of UDP-glucuronic acid to an unknown compound. Overexpression of PsUGT1 sense mRNA has no detectable effect on transgenic pea hairy root cultures or regenerated alfalfa. However, inhibiting PsUGT1 expression by the constitutive expression of antisense mRNA (under the control of the cauliflower mosaic virus 35S promoter) markedly retards growth and development of transgenic alfalfa. Cell structure and organization in the antisense plants are similar to those of controls, but plant growth is reduced and development is delayed. This inhibition in growth is correlated with a twofold delay in the time required for completion of a cell cycle and with a Ͼ 99% inhibition of border cell production. Inhibition of PsUGT1 expression by meristem-localized inducible expression of PsUGT1 antisense mRNA (under the control of its own promoter) is lethal both in pea hairy roots and in transgenic alfalfa plants. These results indicate that PsUGT1 expression is required for normal plant growth and development, and they are consistent with the hypothesis that this UDP-glycosyltransferase regulates activity of a ligand(s) needed for cell division.

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Section: Dna Synthesis In 35s-psugt1 Antisense Plants Is Retarded Commentioning

confidence: 99%

Section: Plant Growthmentioning

confidence: 99%

Meristem-Localized Inducible Expression of a UDP-Glycosyltransferase Gene Is Essential for Growth and Development in Pea and Alfalfa

et al. 1999

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“…and maintain it at about zero for 10 to 12 hours (Jakob andTrosko 1965, Rudolph andDavidson 1975). In V. faba roots, the in itial fall in M. I. is due to the 5-AU blockade of fast cycling cells (Socher andDavidson 1970, 1971) and their cell cycle duration is extended by 7 to 9 hours , even by short treatments with 5-AU (Jakob andTrosko 1965 , Rudolph andDavidson 1975). Cells exposed to 5-AU during Gl however , are delayed by only I to 2 hours.…”

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Recovery from the 5-AU induced blockage in interphase: Evidence for differential recovery.

Davidson

1982

CYTOLOGIA

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Treatment with 5-aminouracil (5-AU) has two effects on proliferating cells: 1) it induces a temporary block at the S-G2 transition or at the end of S phase (Diez, Gonzalez-Fernandez and Lopez-Saez 1976, Scheuermann and Klaffke-Lobsien 1973, Socher and Davidson 1971 and 2) it reduces the rate of DNA synthesis (Jakob and Trosko 1965, Scheuermann andKlaffke-Lobsien 1973). The results of treatment are, first, a fall in mitotic index (MI) and, subsequently, partial synchronization of cells entering mitosis (refs. in Davidson, Golding and Armstrong 1979).The response to 5-AU is as rapid in root primordia of V. faba as in primary roots and fully emerged laterals. This shows that: 1) there is no barrier to the movement of 5-AU through the root cortex and into the primordia (Rudolph and Davidson 1975, Socher andDavidson 1971); 2) the initial response to 5-AU is the same in all meristems of a root system. The rate of recovery from 5-AU, however, as shown by the re-establishment of mitotic activity, is faster in primordia than in primary or lateral roots (Rudolph and Davidson 1975); it is also faster in some cells of a meristem than in other cells (Clowes 1965, Mattingly 1966.One problem in analysing the overall response to 5-AU, i.e., both inhibition and recovery, is that meristems are asynchronous; their cells are distributed through out G1, S, G2 and mitosis. Thus, it is difficult to estimate the extent of the inhibi tion imposed by 5-AU on cells at different stages of interphase because, in normal meristems, cells in these stages cannot be identified. Because of this difficulty we cannot estimate the number of cycles completed by a cell in the presence of 5-AU. One approach to these problems is to follow the behaviour of a marked subpopula tion of cells. Mattingly (1966) induced tetraploid cell formation, with colchicine, treated with 5-AU and showed that the tetraploid cells underwent a complete cell cycle in the presence of 5-AU. Furthermore, these tetraploid cells entered mitosis at the same time as the first diploid cells to recover from the 5-AU blockade.The sensitivity of G, cells to 5-AU has been studied here by treating with col chicine, to induce tetraploid cells, and then treating these cells for two hours during G, with 5-AU. Other root systems were treated with colchicine and then with 5-AU for up to 26 hours. Recovery from 5-AU treatment occurred more rapidly in pri mordia than in primary roots or fully emerged lateral roots, even with a continuous exposure to 5-AU. These results, reported here, suggest that the ability to overcome the block imposed by 5-AU is achieved by whole meristems, not merely by individual cells acting independently. In turn, this suggests a change in the physiological state of a meristem that involves cell-cell interations. This conclusion is largely

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“…In plant roots, a partial synchronization of cell division can be obtained by ex posing the material to an inhibitor of DNA synthesis such as OH-urea (Brulfert and Deysson 1973) or 5-amino uracile (Jacob and Trosko 1965, Mattingly 1966, Prensky and Smith 1965, Wagenaar 1966, but the kinetic parameters of the syn chronous cell populations thus obtained are modified and these populations are made up of cells whose metabolism is disturbed; synchronous cell populations thus obtained have modified kinetic parameters, and are made up of cells whose metabo lism is disturbed; moreover, no single synchronizing agent allows experimentation on all the phases of the mitotic cycle.…”

mentioning

confidence: 99%

Methyl-3 hydroxy-6 quinazoline dione-2-4, a new compound for labelling plant cells. Application to the study of two inhibitors of cytokinesis.

Deysson¹,

Lasselain²,

Pareyre³

1980

CYTOLOGIA

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The relation between 5-amino uracil-induced mitotic synchronization and DNA synthesis

Cited by 39 publications

References 8 publications

Meristem-Localized Inducible Expression of a UDP-Glycosyltransferase Gene Is Essential for Growth and Development in Pea and Alfalfa

Meristem-Localized Inducible Expression of a UDP-Glycosyltransferase Gene Is Essential for Growth and Development in Pea and Alfalfa

Recovery from the 5-AU induced blockage in interphase: Evidence for differential recovery.

Methyl-3 hydroxy-6 quinazoline dione-2-4, a new compound for labelling plant cells. Application to the study of two inhibitors of cytokinesis.

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