Abstract
Progression through G2/M of the mammalian cell division cycle requires the coordinated expression of many gene products, but little is known of the transcriptional regulators involved. Schizosaccharomyces pombe Cdc5 is a putative transcription factor implicated in G2/M transit. We recently identified a cDNA encoding a putative human transcription factor, now designated human Cdc5 (hCdc5), with homology to S. pombe Cdc5. Widespread expression of hCdc5 in human tissues and homology with expressed sequences in other eukaryotes suggested an evolutionarily conserved general function. Nuclear import of hCdc5 upon serum stimulation of mammalian cells suggested a possible role in cell proliferation. We now report that overexpression of hCdc5 in mammalian cells shortened G2 and reduced cell size. A dominant negative mutant of hCdc5 lacking the carboxyl-terminal activation domain slowed G2 progression and delayed entry into mitosis. Thus, hCdc5 is the first transcriptional regulator shown to affect G2 progression and mitotic entry in mammalian cells.
Highlights
Orchestration of the cell division cycle includes a series of checkpoints which ensure that some events are completed before others begin [1]
We recently identified a cDNA encoding a putative human transcription factor [20] with homology to S. pombe Cdc5 [19]
A dominant negative mutant of human Cdc5 (hCdc5) lacking the carboxyl-terminal activation domain slowed G2 progression and delayed entry into mitosis. These results demonstrate that hCdc5 regulates G2 progression and mitotic entry and is the first transcriptional regulator shown to effect G2/M transit in mammalian cells
Summary
Orchestration of the cell division cycle includes a series of checkpoints which ensure that some events are completed before others begin [1]. These results demonstrate that hCdc5 regulates G2 progression and mitotic entry and is the first transcriptional regulator shown to effect G2/M transit in mammalian cells. Cell line 3T15.8 was generated by transfecting BALBc/3T3 cells with pUHD 15–1 (Display Systems Biotechnology), which directs the expression of a tetracycline repressor-VP16 fusion protein, selecting in the presence of 400 g/ml G418.
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