Identification of G1-regulated genes in normally cycling human cells

Maroun J. Beyrouthy, Karen E. Alexander, Amy Baldwin, Michael L. Whitfield, Hank W. Bass, Dan McGee, Myra M. Hurt

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: Obtaining synchronous cell populations is essential for cell-cycle studies. Methods such as serum withdrawal or use of drugs which block cells at specific points in the cell cycle alter cellular events upon re-entry into the cell cycle. Regulatory events occurring in early G1 phase of a new cell cycle could have been overlooked. Methodology and Findings: We used a robotic mitotic shake-off apparatus to select cells in late mitosis for genome-wide gene expression studies. Two separate microarray experiments were conducted, one which involved isolation of RNA hourly for several hours from synchronous cell populations, and one experiment which examined gene activity every 15 minutes from late telophase of mitosis into G1 phase. To verify synchrony of the cell populations under study, we utilized methods including BrdU uptake, FACS, and microarray analyses of histone gene activity. We also examined stress response gene activity. Our analysis enabled identification of 200 early G1-regulated genes, many of which currently have unknown functions. We also confirmed the expression of a set of genes candidates (fos, atf3 and tceb) by qPCR to further validate the newly identified genes. Conclusion and Significance: Genome-scale expression analyses of the first two hours of G1 in naturally cycling cells enabled the discovery of a unique set of G1-regulated genes, many of which currently have unknown functions, in cells progressing normally through the cell division cycle. This group of genes may contain future targets for drug development and treatment of human disease.

Original languageEnglish (US)
Article numbere3943
JournalPLoS One
Volume3
Issue number12
DOIs
StatePublished - Dec 15 2008
Externally publishedYes

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Genes
Cells
Cell Cycle
cell cycle
genes
cells
G1 Phase
Mitosis
interphase
mitosis
Microarrays
Genome
Population
Telophase
fos Genes
Spindle Apparatus
drugs
telophase
Robotics
Human Development

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Beyrouthy, M. J., Alexander, K. E., Baldwin, A., Whitfield, M. L., Bass, H. W., McGee, D., & Hurt, M. M. (2008). Identification of G1-regulated genes in normally cycling human cells. PLoS One, 3(12), [e3943]. https://doi.org/10.1371/journal.pone.0003943

Identification of G1-regulated genes in normally cycling human cells. / Beyrouthy, Maroun J.; Alexander, Karen E.; Baldwin, Amy; Whitfield, Michael L.; Bass, Hank W.; McGee, Dan; Hurt, Myra M.

In: PLoS One, Vol. 3, No. 12, e3943, 15.12.2008.

Research output: Contribution to journalArticle

Beyrouthy, MJ, Alexander, KE, Baldwin, A, Whitfield, ML, Bass, HW, McGee, D & Hurt, MM 2008, 'Identification of G1-regulated genes in normally cycling human cells', PLoS One, vol. 3, no. 12, e3943. https://doi.org/10.1371/journal.pone.0003943
Beyrouthy MJ, Alexander KE, Baldwin A, Whitfield ML, Bass HW, McGee D et al. Identification of G1-regulated genes in normally cycling human cells. PLoS One. 2008 Dec 15;3(12). e3943. https://doi.org/10.1371/journal.pone.0003943
Beyrouthy, Maroun J. ; Alexander, Karen E. ; Baldwin, Amy ; Whitfield, Michael L. ; Bass, Hank W. ; McGee, Dan ; Hurt, Myra M. / Identification of G1-regulated genes in normally cycling human cells. In: PLoS One. 2008 ; Vol. 3, No. 12.
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