Cell size checkpoint control by the retinoblastoma tumor suppressor pathway.
Size control is essential for all proliferating cells, and is thought to be regulated by checkpoints that couple cell size to cell cycle progression. The aberrant cell-size phenotypes caused by mutations in the retinoblastoma (RB) tumor suppressor pathway are consistent with a role in size checkpoint control, but indirect effects on size caused by altered cell cycle kinetics are difficult to rule out. The multiple fission cell cycle of the unicellular alga Chlamydomonas reinhardtii uncouples growth from division, allowing direct assessment of the relationship between size phenotypes and checkpoint function. Mutations in the C. reinhardtii RB homolog encoded by MAT3 cause supernumerous cell divisions and small cells, suggesting a role for MAT3 in size control. We identified suppressors of an mat3 null allele that had recessive mutations in DP1 or dominant mutations in E2F1, loci encoding homologs of a heterodimeric transcription factor that is targeted by RB-related proteins. Significantly, we determined that the dp1 and e2f1 phenotypes were caused by defects in size checkpoint control and were not due to a lengthened cell cycle. Despite their cell division defects, mat3, dp1, and e2f1 mutants showed almost no changes in periodic transcription of genes induced during S phase and mitosis, many of which are conserved targets of the RB pathway. Conversely, we found that regulation of cell size was unaffected when S phase and mitotic transcription were inhibited. Our data provide direct evidence that the RB pathway mediates cell size checkpoint control and suggest that such control is not directly coupled to the magnitude of periodic cell cycle transcription.
Fang
Su-Chiung
SC
de los Reyes
Chris
C
Umen
James G
JG
D000546:Amanitins / Q000494:pharmacology; D000818:Animals; D002453:Cell Cycle / Q000502:physiology*; D048429:Cell Size; D016825:Chlamydomonas reinhardtii / Q000166:cytology* / Q000378:metabolism*; D050687:E2F1 Transcription Factor / Q000378:metabolism; D005799:Genes, Dominant; D005816:Genetic Complementation Test; D007700:Kinetics; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D010641:Phenotype; D015800:Protozoan Proteins / Q000302:isolation & purification / Q000378:metabolism*; D012319:RNA Polymerase II / Q000037:antagonists & inhibitors; D016160:Retinoblastoma Protein / Q000302:isolation & purification / Q000378:metabolism*; D013489:Suppression, Genetic; D050680:Transcription Factor DP1 / Q000378:metabolism; D014158:Transcription, Genetic
D000546:Amanitins; D050687:E2F1 Transcription Factor; D015800:Protozoan Proteins; D016160:Retinoblastoma Protein; D050680:Transcription Factor DP1; C432865:mat3 protein, Chlamydomonas; D012319:RNA Polymerase II