Expression analysis using DNA microarrays demonstrates that E2F-1 up-regulates expression of DNA replication genes including replication protein A2.
The transcription factor E2F-1 plays a pivotal role in the regulation of G1/S transition in higher eukaryotes cell cycle. We used a cell line containing an inducible E2F-1 and oligonucleotide microarray analysis to identify novel E2F target genes. We show that E2F-1 up-regulates the expression of a number of genes coding for components of the DNA replication machinery. Among them is the gene coding for the 32 Kd subunit of replication protein A (RPA2). Replication protein A is the most abundant single strand DNA binding complex and it is essential for DNA replication. We demonstrate that RPA2 is a novel E2F target gene whose expression can be directly regulated by E2F-1 via E2F binding sites in its promoter. In addition, expression of Topoisomerase IIalpha and subunit IV of DNA polymerase alpha is also up-regulated upon E2F-1 induction. Taken together, these results provide novel links between components of the DNA replication machinery and the cell growth regulatory pathway involving the Rb tumor suppressor and E2F.
Kalma
Y
Y
Department of Molecular Cell Biology, The Weizmann Institute of Science, Rehovot 76100, Israel.
Marash
L
L
Lamed
Y
Y
Ginsberg
D
D
D000951:Antigens, Neoplasm; D001483:Base Sequence; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004256:DNA Polymerase I / Q000096:biosynthesis; D004261:DNA Replication / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; D020869:Gene Expression Profiling; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D011401:Promoter Regions, Genetic; D021122:Protein Subunits; D051716:Replication Protein A; D056465:Retinoblastoma-Binding Protein 1; D050680:Transcription Factor DP1; D014157:Transcription Factors / Q000378:metabolism*; D015854:Up-Regulation
D000951:Antigens, Neoplasm; D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D050687:E2F1 Transcription Factor; C493753:E2F1 protein, human; D007527:Isoenzymes; D021122:Protein Subunits; C497167:RPA1 protein, human; D051716:Replication Protein A; D056465:Retinoblastoma-Binding Protein 1; D050680:Transcription Factor DP1; D014157:Transcription Factors; D004256:DNA Polymerase I; D004250:DNA Topoisomerases, Type II