Mammalian cells lack checkpoints for tetraploidy, aberrant centrosome number, and cytokinesis failure.
BACKGROUND
Mammalian cells have been reported to have a p53-dependent tetraploidy checkpoint that blocks cell cycle progression in G1 in response to failure of cell division. In most cases where the tetraploidy checkpoint has been observed cell division was perturbed by anti-cytoskeleton drug treatments. However, other evidence argues against the existence of a tetraploidy checkpoint. Cells that have failed to divide differ from normal cells in having two nuclei, two centrosomes, a decreased surface to volume ratio, and having undergone an abortive cytokinesis. We tested each of these to determine which, if any, cause a G1 cell cycle arrest.


RESULTS
Primary human diploid fibroblasts with intact cell cycle checkpoints were used in all experiments. Synchronized cells exhibited G1 arrest in response to division failure caused by treatment with either cytochalasin or the myosin II inhibitor blebbistatin. The role of tetraploidy, aberrant centrosome number, and increased cell size were tested by cell/cell and cell/cytoplast fusion experiments; none of these conditions resulted in G1 arrest. Instead we found that various drug treatments of the cells resulted in cellular damage, which was the likely cause of the arrest. When cytokinesis was blocked in the absence of damage-inducing drug treatments no G1 arrest was observed.


CONCLUSIONS
We show that neither tetraploidy, aberrant centrosome number, cell size, nor failure of cytokinesis lead to G1 arrest, suggesting that there is no tetraploidy checkpoint. Rather, certain standard synchronization treatments cause damage that is the likely cause of G1 arrest. Since tetraploid cells can cycle when created with minimal manipulation, previous reports of a tetraploidy checkpoint can probably be explained by side effects of the drug treatments used to observe them.
Wong
Connie
C
Department of Biological Sciences, Stanford University, Stanford, CA 94305, USA. conniewg@stanford.edu
Stearns
Tim
T
D002453:Cell Cycle / Q000187:drug effects* / Q000235:genetics*; D048430:Cell Shape; D018385:Centrosome; D003572:Cytochalasins / Q000494:pharmacology; D048749:Cytokinesis; D005347:Fibroblasts / Q000166:cytology; D016193:G1 Phase / Q000187:drug effects; D006576:Heterocyclic Compounds, 4 or More Rings / Q000494:pharmacology; D006801:Humans; D011123:Polyploidy
D003572:Cytochalasins; D006576:Heterocyclic Compounds, 4 or More Rings; C472645:blebbistatin