Isolation and characterization of the E2F-like gene in plants.
The transcription factor E2F regulates the expression of genes involved in the progression of G1/S transition and DNA replication in mammalian cells. We cloned and characterized a cDNA (NtE2F) corresponding to a E2F homolog of tobacco (Nicotiana tabacum). The transcription of NtE2F was induced as cells progressed from G1 to the S phase and expressed much earlier than that of the proliferating cell nuclear antigen (PCNA) gene. We demonstrated that NtE2F can interact with the tobacco retinoblastoma (Rb)-related protein in a yeast two-hybrid assay. To further characterize NtE2F, the trans-activation activity of NtE2F was examined by using a transient assay in the tobacco Bright Yellow-2 (BY-2) cells with NtE2F fused to the DNA-binding domain of the veast transcriptional activator GAL4. NtE2F activated the transcription of the beta-glucuronidase (GUS) reporter gene driven by a cauliflower mosaic virus (CaMV) 35S core promoter containing the GAL4-binding sequence. This is the first report of the identification of a functionally equivalent E2F-like gene in plants.
Sekine
M
M
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan. sekine@bs.aist-nara.ac.jp
Ito
M
M
Uemukai
K
K
Maeda
Y
Y
Nakagami
H
H
Shinmyo
A
A
D000595:Amino Acid Sequence; D002352:Carrier Proteins; D018797:Cell Cycle Proteins / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D016160:Retinoblastoma Protein / Q000235:genetics; D056465:Retinoblastoma-Binding Protein 1; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D014026:Nicotiana; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*; D015003:Yeasts
D002352:Carrier Proteins; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D050684:E2F Transcription Factors; D018809:Proliferating Cell Nuclear Antigen; D012333:RNA, Messenger; D016160:Retinoblastoma Protein; D056465:Retinoblastoma-Binding Protein 1; D014157:Transcription Factors