Related Experiment Videos
Cloning, chromosomal location, and characterization of mouse E2F1
1McArdle Laboratory for Cancer Research, University of Wisconsin, Madison 53706.
Molecular and Cellular Biology
|March 1, 1994
Summary
Mouse E2F1, a protein similar to human E2F1, was cloned and found to activate growth-regulated promoters. Its mRNA levels correlate with cell cycle progression, suggesting a role in growth factor signaling.
Area of Science:
- Molecular Biology
- Cell Cycle Regulation
- Gene Expression
Background:
- E2F proteins are crucial for cell growth control, binding to retinoblastoma protein and E2F sites in growth-regulated genes.
- Previous studies identified distinct human E2F1 and mouse DP-1 proteins, differing significantly in sequence.
Purpose of the Study:
- To clone and characterize mouse E2F1, a homolog of human E2F1.
- To investigate the role of E2F1 and DP-1 in mouse NIH 3T3 cell cycle progression.
- To determine E2F1's ability to transactivate growth-regulated promoters.
Main Methods:
- Cloning of mouse E2F1 cDNA and characterization of its encoded protein.
- Mapping of the mouse E2F1 gene to chromosome 2 using specific primers.
- Analysis of E2F1 and DP-1 mRNA levels during different cell cycle stages.
- Assessment of E2F1's transactivation potential on various promoters (dhfr, thymidine kinase, DNA polymerase alpha, thymidylate synthase, cad, c-myc).
Main Results:
- A mouse cDNA encoding a protein 86% identical to human E2F1 was successfully cloned.
- Mouse E2F1 gene localized to chromosome 2.
- E2F1 mRNA levels, but not DP-1, correlated with changes in dhfr promoter transcription during the cell cycle.
- E2F1 demonstrated transactivation of dhfr, thymidine kinase, and DNA polymerase alpha promoters, but not others.
Conclusions:
- Mouse E2F1 is a functional homolog of human E2F1.
- E2F1 plays a significant role in regulating cell cycle-dependent gene expression.
- E2F1 likely mediates growth factor-initiated signal transduction pathways.