|
| Status |
Public on Apr 01, 2010 |
| Title |
Global analysis of differential polyadenylation of mRNAs during mitotic cell cycle: S phase versus G2/M phase. |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
|
| Summary |
We compared the poly(A) tail length status of mRNAs of HeLa cells between two phases of the mitotic cell cycle: S and G2/M phases. Hundreds of mRNAs were found to be regulated by changes in their poly(A) tail length during mitotic cell cycle in a phase specific manner. Many of these differentially polyadenylated mRNAs encode proteins related to cell death, cell cycle and cellular growth and proliferation. HeLa cells were synchronized with double thymidine blockade (12 hours with 2 mM thymidine, 12 hours release, and 12 hours with 2 mM thymidine), and samples were taken after 2 hours release (S phase) and 8 hours release (G2/M phase). For each condition total RNA was purified by two different procedures: poly(U) chromatography and oligo(dT)-chromatography. Poly(U)-chromatography (Jacobson, 1987): 100 μg of total RNA were bound to poly(U)-sepharose (Sigma) and eluted at 35ºC to isolate mRNAs with short poly(A) tail (<30As, SHORT fraction). Oligo(dT) chromatography: mRNAs were purified independently of their poly(A) tail length with Ambion Poly(A)Purist kit from 20 μg total RNA (ALL fraction). Jacobson, A. Purification and fractionation of poly(A)+ RNA. Methods in Enzymology (1987) 152: 254-261.
Keywords: time course
|
| |
|
| Overall design |
Comparison of ALL fraction mRNAs and SHORT fraction mRNAs measured after 2 hours (S phase) and 8 hours release (G2/M) from double thymidine blockade; 3 biological replicates at each of the two time points; two technical replicates with dye swapping per comparison.
|
| |
|
| Contributor(s) |
Novoa I, Gallego J, Mendez R |
| Citation(s) |
20364142 |
| Submission date |
Mar 25, 2009 |
| Last update date |
Dec 06, 2012 |
| Contact name |
Isabel Novoa |
| E-mail(s) |
isabel.novoa@crg.es
|
| Organization name |
Center for Genomic Regulation
|
| Department |
Gene Regulation
|
| Street address |
Dr. Aiguader, 88
|
| City |
Barcelona |
| ZIP/Postal code |
08003 |
| Country |
Spain |
| |
|
| Platforms (1) |
| GPL887 |
Agilent-012097 Human 1A Microarray (V2) G4110B (Feature Number version) |
|
| Samples (12)
|
| GSM386473 |
ALL fraction mRNAs 2 hours versus SHORT fraction mRNAs 2 hours replicate 1 |
| GSM386474 |
ALL fraction mRNAs 2 hours versus SHORT fraction mRNAs 2 hours replicate 1 dye-swap |
| GSM386475 |
ALL fraction mRNAs 2 hours versus SHORT fraction mRNAs 2 hours replicate 2 |
| GSM386476 |
ALL fraction mRNAs 2 hours versus SHORT fraction mRNAs 2 hours replicate 2 dye-swap |
| GSM386477 |
ALL fraction mRNAs 2 hours versus SHORT fraction mRNAs 2 hours replicate 3 |
| GSM386478 |
ALL fraction mRNAs 2 hours versus SHORT fraction mRNAs 2 hours replicate 3 dye-swap |
| GSM386479 |
ALL fraction mRNAs 8 hours versus SHORT fraction mRNAs 8 hours replicate 1 |
| GSM386480 |
ALL fraction mRNAs 8 hours versus SHORT fraction mRNAs 8 hours replicate 1 dye-swap |
| GSM386481 |
ALL fraction mRNAs 8 hours versus SHORT fraction mRNAs 8 hours replicate 2 |
| GSM386482 |
ALL fraction mRNAs 8 hours versus SHORT fraction mRNAs 8 hours replicate 2 dye-swap |
| GSM386483 |
ALL fraction mRNAs 8 hours versus SHORT fraction mRNAs 8 hours replicate 3 |
| GSM386484 |
ALL fraction mRNAs 8 hours versus SHORT fraction mRNAs 8 hours replicate 3 dye-swap |
|
| Relations |
| BioProject |
PRJNA115881 |
Less...
More...