Abstract

Under optimal growing conditions, synchronous cultures of the alga Scenedesmus quadricauda underwent three DNA replications and three mitoses during one cell cycle. This resulted in eight daughter cells. By different illumination regimes and temporal addition of cycloheximide, cell divisions resulting in two, four or eight daughter cells per cycle were obtained. The selected cell cycle patterns differed in timing of commitment points, the number of nuclear divisions, and their positioning in the cell cycle. These distinct cell cycle patterns allowed to assess the correlation of histone H1 kinase activity with commitment points and mitoses. The activity of the histone H1 kinases was assayed in cellular protein extracts and after affinity purification using the p13 suc1 protein. The main peaks of kinase activity in the cellular extract were found to correlate with the commitment points. Small histone H1 kinase activity peaks were also found which preceded the nuclear division. Contrary to the histone H1 kinase activity of cellular extracts, the p13 suc1-bound kinase activity preceded the nuclear division, whilst its activity was negligible at the commitment points. Being able to manipulate the timing of commitment points and cell division by manipulating experimental conditions, we could precisely match the commitment points to an as yet unidentified histone H1 kinase activity and mitosis to p13 suc1-bound CDK activity during a particular cell division pattern with overlapping cycles. This provides molecular evidence, that local activation of CDKs regulates distinct events of the cell cycle.

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