Abstract
Although sucrose availability is crucial for commitment to plant cell division during G1 phase by controlling the expression of D-type cyclins, it has remained unclear how these factors mediate entry into the cell cycle. Here we show that Arabidopsis RETINOBLASTOMA-RELATED PROTEIN 1 (AtRBR1) is involved in G1-phase cell cycle arrest caused by sucrose starvation. We generated estrogen-inducible AtRBR1 RNA interference (RNAi) Arabidopsis suspension MM2d cells, and found that downregulation of AtRBR1 leads to a higher frequency of arrest in G2 phase, instead of G1-phase arrest in the uninduced control, after sucrose starvation. Synchronization experiments confirmed that downregulation of AtRBR1 leads to a prolonged G2 phase and delayed activation of G2/M marker genes. Downregulation of AtRBR1 also stimulated the activation of E2F-regulated genes when these genes were repressed in the uninduced cells under the limited sucrose conditions. We conclude that AtRBR1 is a key effector for the ability of sucrose to modulate progression from G1 phase.
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Abbreviations
- ACT:
-
Actin
- ChIP:
-
Chromatin immunoprecipitation
- cDNA:
-
Complementary DNA
- CDC:
-
Cell division control
- CDK:
-
Cyclin-dependent kinase
- DP:
-
DNA-binding heterodimerization partner protein
- E2F:
-
Adenovirus E2 promoter-binding factor
- GST:
-
Glutatione S-transferase
- HA:
-
Hemagglutinin
- IP:
-
Immunoprecipitation
- LSC:
-
Laser scanning cytometer
- ORF:
-
Open reading frame
- MBP:
-
Maltose-binding protein
- MCM:
-
Mini chromosome maintenance
- MCS:
-
Multi-cloning site
- PCR:
-
Polymerase chain reaction
- PCNA:
-
Proliferating cell nuclear antigen
- RB:
-
Retinoblastoma
- RBR:
-
Retinoblastoma-related
- RNAi:
-
RNA interference
- RNR:
-
Ribonucleotide reductase
- RT-PCR:
-
Reverse transcriptase polymerase chain reaction
- WCE:
-
Whole cell extract
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Acknowledgements
The authors wish to thank Drs. Ko Kato and Kazuya Yoshida for their helpful discussions and suggestions throughout this work, and Dr. Ian Smith for critical reading of the manuscript. We also thank Dr. Masaaki Umeda for providing access to a flow cytometer. This research was supported by a Grant-in-Aid for Scientific Research from the Japanese Ministry of Education, Culture, Sports, Science and Technology and by a Grant-in-Aid for Creative Scientific Research.
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Hirano, H., Harashima, H., Shinmyo, A. et al. Arabidopsis RETINOBLASTOMA-RELATED PROTEIN 1 is involved in G1 phase cell cycle arrest caused by sucrose starvation. Plant Mol Biol 66, 259–275 (2008). https://doi.org/10.1007/s11103-007-9268-2
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DOI: https://doi.org/10.1007/s11103-007-9268-2