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Cell cycle-dependent gene expression in V point-arrested BALB/c-3T3 cells
J E Olson1, J T Winston, J A Whitlock
1Department of Cell Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232.
Journal of Cellular Physiology
|February 1, 1993
Summary
Density-arrested BALB/c-3T3 cells were used to study gene expression during the G1 phase of the cell cycle. This method identified distinct early and late G1 gene expression patterns, revealing c-myb
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- BALB/c-3T3 cells arrest in mid-G1 (V point) when stimulated to proliferate in amino acid-deficient medium.
- Release from V point arrest requires 6 hours to enter S phase, with mRNA synthesis crucial for the initial 2-3 hours.
Purpose of the Study:
- To analyze gene expression during late G1 using a mid-G1 arrest protocol.
- To investigate the role of c-myb in late G1 progression and DNA synthesis.
Main Methods:
- Utilizing a mid-G1 arrest protocol in BALB/c-3T3 cells.
- Analyzing gene expression patterns before and after release from V point arrest.
- Employing c-myb antisense oligonucleotides to assess gene function.
Main Results:
- Early G1 gene induction was unaffected, but late G1 gene expression was delayed until after V point release.
- Late G1 gene expression timing post-release mirrored that of quiescent G0 cell stimulation.
- c-myb expression in late G1 was found to be essential for traversing late G1 and initiating DNA synthesis.
Conclusions:
- The V point arrest protocol effectively categorizes gene expression into early (pre-V point) and late (post-V point) phases.
- This method is valuable for studying growth factor-regulated events controlling late G1 progression and DNA synthesis commitment.
- Late G1 c-myb expression is a critical requirement for BALB/c-3T3 fibroblasts to proceed through late G1 and initiate DNA synthesis.