Analysis of variation of amplitudes in cell cycle gene expression.
BACKGROUND
Variation in gene expression among cells in a population is often considered as noise produced from gene transcription and post-transcription processes and experimental artifacts. Most studies on noise in gene expression have emphasized a few well-characterized genes and proteins. We investigated whether different cell-arresting methods have impacts on the maximum expression levels (amplitudes) of a cell cycle related gene.


RESULTS
By introducing random noise, modeled by a von Mises distribution, to the phase angle in a sinusoidal model in a cell population, we derived a relationship between amplitude and the distribution of noise in maximum transcription time (phase). We applied our analysis to Whitfield's HeLa cell cycle data. Our analysis suggests that among 47 cell cycle related genes common to the 2nd experiment (thymidine-thymidine method) and the 4th experiment (thymidine-nocodazole method): (i) the amplitudes of CDC6 and PCNA, which are expressed during G1/S phase, are smaller in the 2nd experiment than in the 4th, while the amplitude of CDC20, which is expressed during G2/M phase, is smaller in the 4th experiment; and (ii) the two cell-arresting methods had little impact on the amplitudes of the other 43 genes in the 2nd and 4th experiments.


CONCLUSIONS
Our analysis suggests that procedures that arrest cells in different stages of the cell cycle differentially affect expression of some cell cycle related genes once the cells are released from arrest. The impact of the cell-arresting method on expression of a cell cycle related gene can be quantitatively estimated from the ratio of two estimated amplitudes in two experiments. The ratio can be used to gauge the variation in the phase/peak expression time distribution involved in stochastic transcription and post-transcriptional processes for the gene. Further investigations are needed using normal, unperturbed and synchronized HeLa cells as a reference to compare how many cell cycle related genes are directly and indirectly affected by various cell-arresting methods.
Liu
Delong
D
CIIT Ceters for Health Research, 6 Davis Drive, Research Triangle Park, NC 27709, USA. dliu@ciit.org
Gaido
Kevin W
KW
Wolfinger
Russ
R
D064199:Cdc20 Proteins; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics*; D018797:Cell Cycle Proteins / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D008954:Models, Biological; D015739:Nocodazole / Q000494:pharmacology; D009687:Nuclear Proteins / Q000235:genetics; D018809:Proliferating Cell Nuclear Antigen / Q000235:genetics; D013936:Thymidine
C103706:CDC6 protein, human; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; C577488:CDC20 protein, human; D015739:Nocodazole; D013936:Thymidine