Inhibition of serine/threonine-specific protein phosphatases causes premature activation of cdc2MsF kinase at G2/M transition and early mitotic microtubule organisation in alfalfa.
Reversible phosphorylation of serine/threonine residues of cell cycle-regulatory proteins is one of the key molecular mechanisms controlling eukaryotic cell division. In plants, the protein kinase partners (i.e. p34cdc2/CDC28-related kinases) have been extensively studied, while the role of counter-acting protein phosphatases is less well understood. We used endothall (ET) as a cell-permeable inhibitor of serine/threonine-specific protein phosphatases to alter cytological and biochemical characteristics of cell division in cultured alfalfa cells. A high concentration of ET (10 and 50 microM) inhibited both protein phosphatases 1 and 2 (PP1 and PP2A), while a low concentration (1 microM) of ET-treatment primarily reduced the PP2A activity. High concentrations of the inhibitor increased the frequency of hypercondensed early and late prophase chromosomes that could not enter metaphase. In contrast, a low concentration of ET did not interfere with chromosomal events but caused significant alterations in the organisation of microtubules. Exposure of cells to 1 microM ET resulted in disturbance of preprophase band formation, increase in the number of nuclei with prophase microtubule assembly, premature polarisation of the spindle, and abnormal phragmoplast maturation. Under the same conditions, the ET-treated cells exhibited an early increase in cdc2MsF kinase activity. These results suggest that PP2A contributes to the control of mitotic kinase activities and microtubule organisation. Normal chromosome condensation and mitotic progression are dependent on both PP1 and PP2A activities. The presented data support the functional role of protein phosphatases in the co-ordination of chromosomal and microtubule events in dividing plant cells.
Institute of Plant Biology, Biological Research Center, Hungarian Academy of Sciences, H-6701 Szeged, PO Box 521, Hungary.
F
F
Ayaydin
E
E
Vissi
T
T
Mészáros
P
P
Miskolczi
I
I
Kovács
A
A
Fehér
V
V
Dombrádi
F
F
Erdödi
P
P
Gergely
D
D
Dudits
D002875:Chromosomes; D003998:Dicarboxylic Acids / Q000494:pharmacology; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016195:G2 Phase; D000455:Medicago sativa / Q000201:enzymology* / Q000648:ultrastructure; D008870:Microtubules / Q000648:ultrastructure*; D008938:Mitosis; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*
D003998:Dicarboxylic Acids; D004791:Enzyme Inhibitors; C100030:endothall; D010749:Phosphoprotein Phosphatases