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Figure 1


Fig. 1. Cell-cycle-dependent phosphorylation pattern of AtMAP65-1. (A) Cell cycle synchronisation. Graphs show the frequency of metaphase (open diamonds), anaphase (open squares) and telophase (open triangles) cells in the synchronised BY-2 cell culture. Eleven sampling points were analysed: points 0-2 were collected every two hours after aphidicolin was washed out, points 3-5 were collected every 2 hours during propyzamide treatment and points 6-10 were collected every 40 minutes after propyzamide was washed out with the exception that there was a 2-hour interval between points 9 and 10. (B) Autoradiogram (i) and Coomassie Blue-stained gel (ii) showing phosphorylation of AtMAP65-1 using the total protein extract from cells collected at sampling points 0-10 described in A (i). 5 µg of recombinant AtMAP65-1 was used in each assay. The Coomassie Blue-stained gel of the extracts used as kinase in (i) and (ii) is shown in (iii). (C) Two-dimensional SDS-PAGE immunoblots of total protein extracts from interphase cells (S-phase, sampling point 0), metaphase cells (M-phase, sampling point 4) and M-phase extract treated with phosphatase, probed with anti AtMAP65-1. The area of the membrane where AtMAP65-1 isoforms were detected is shown.





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