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Molecular and Cellular Biology, July 2001, p. 4684-4699, Vol. 21, No. 14
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.14.4684-4699.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Role for E2F in Control of Both DNA Replication and Mitotic Functions as Revealed from DNA Microarray Analysis

Seiichi Ishida,1 Erich Huang,1 Harry Zuzan,2 Rainer Spang,2 Gustavo Leone,1,dagger Mike West,2 and Joseph R. Nevins1,*

Department of Genetics and Howard Hughes Medical Institute, Duke University Medical Center, Durham, North Carolina 277101 and Institute of Statistics and Decision Sciences, Duke University, Durham, North Carolina 277082

Received 8 January 2001/Returned for modification 1 February 2001/Accepted 26 April 2001

We have used high-density DNA microarrays to provide an analysis of gene regulation during the mammalian cell cycle and the role of E2F in this process. Cell cycle analysis was facilitated by a combined examination of gene control in serum-stimulated fibroblasts and cells synchronized at G1/S by hydroxyurea block that were then released to proceed through the cell cycle. The latter approach (G1/S synchronization) is critical for rigorously maintaining cell synchrony for unambiguous analysis of gene regulation in later stages of the cell cycle. Analysis of these samples identified seven distinct clusters of genes that exhibit unique patterns of expression. Genes tend to cluster within these groups based on common function and the time during the cell cycle that the activity is required. Placed in this context, the analysis of genes induced by E2F proteins identified genes or expressed sequence tags not previously described as regulated by E2F proteins; surprisingly, many of these encode proteins known to function during mitosis. A comparison of the E2F-induced genes with the patterns of cell growth-regulated gene expression revealed that virtually all of the E2F-induced genes are found in only two of the cell cycle clusters; one group was regulated at G1/S, and the second group, which included the mitotic activities, was regulated at G2. The activation of the G2 genes suggests a broader role for E2F in the control of both DNA replication and mitotic activities.

* Corresponding author. Mailing address: Department of Genetics, Duke University Medical Center, Durham, NC 27710. Phone: (919) 684-2746. Fax: (919) 681-8973. E-mail: J.Nevins{at}duke.edu.

dagger Present address: Division of Human Cancer Genetics, Ohio State University, Columbus, OH 43210.

Molecular and Cellular Biology, July 2001, p. 4684-4699, Vol. 21, No. 14
0270-7306/01/$04.00+0   DOI: 10.1128/MCB.21.14.4684-4699.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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