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L-Mimosine (beta-N-[3-hydroxy-4-pyridone]-alpha-aminopropionic acid)--a rare amino acid derived from Mimosa and Leucaena plants--arrests cells reversibly late during G1 phase or at the beginning of S-phase. If mimosine were to arrest cells immediately before S-phase, it would provide a superb tool for the investigation of the initiation of DNA synthesis. Therefore, we reexamined the point of action of mimosine. Mitotic HeLa cells were released into 200 microM mimosine and grown for approximately 10 h to block them, before the cells were permeabilized and the amino acid removed by washing them thoroughly. On addition of the appropriate triphosphates, DNA synthesis-measured by the incorporation of [32P]dTTP--began immediately; as it is known that such permeabilized cells cannot initiate DNA synthesis but can only resume elongating previously initiated chains, mimosine must arrest after DNA synthesis has begun. Moreover, cells grown in mimosine assembled functional replication factories--detected by immunolabeling after incorporation of biotin-dUTP--that were typical of those found early during S-phase. Disappointingly, it seems that mimosine--like aphidocolin--blocks only after cells enter S-phase.

Original publication

DOI

10.1006/excr.1996.0035

Type

Journal article

Journal

Exp Cell Res

Publication Date

01/02/1996

Volume

222

Pages

275 - 280

Keywords

Amino Acids, Cell Cycle, DNA, DNA Replication, Dose-Response Relationship, Drug, G1 Phase, HeLa Cells, Humans, Mimosine, Mitosis, S Phase, Time Factors