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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Michelotti, Gregory A. Schwinn, Debra A. Morris, Daniel P. |
| Description | Author Affiliation: Morris DP ( Department of Anesthesiology, Duke University Medical Center, Durham, North Carolina 27710, USA. morri082@mc.duke.edu) |
| Abstract | Using an improved chromatin immunoprecipitation assay designed to increase immunoprecipitation efficiency, we investigated changes in RNA polymerase II (Pol II) density and carboxyl-terminal domain (CTD) phosphorylation during transcription of the cyclophilin A (PPIA), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and several androgen-responsive genes in LNCaP cells. As generally observed in higher eukaryotes, promoter proximal pausing of Pol II appeared to occur on the PPIA and GAPDH genes, but apparently not on the androgen-responsive genes PSA and NKX3-1. Unlike some mammalian studies, we found that the CTD of Pol II in promoter regions contains little phosphorylation at Ser-2 of the heptad repeat, suggesting that Ser-2 phosphorylation is not involved in polymerase exit from the promoter region. In contrast, Pol II near the promoter displayed high levels of Ser-5 phosphorylation, which decreased as polymerase transcribed beyond the promoter region of the PPIA and GAPDH genes. However, total Pol II levels appear to decrease as much or more, suggesting that Ser-5 phosphorylation is maintained. In support of this conclusion, a phosphoserine 5-specific antibody quantitatively immunoprecipitates native hyperphosphorylated Pol II, suggesting that all polymerase with phosphoserine 2 also contains phosphoserine 5. Given reports indicating that phosphoserine 5 is present during elongation in yeast, our data suggest that gross changes in CTD phosphorylation patterns during transcription may be more conserved in yeast and humans than recognized previously. |
| ISSN | 00219258 |
| e-ISSN | 1083351X |
| Journal | Journal of Biological Chemistry |
| Issue Number | 36 |
| Volume Number | 280 |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology (United States) |
| Publisher Date | 2005-09-09 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | RNA Polymerase II Metabolism Repetitive Sequences, Amino Acid Saccharomyces Cerevisiae Enzymology Androgens Physiology Animals Cell Line Conserved Sequence Cyclophilin A Biosynthesis Genetics Formaldehyde Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) Phosphorylation Rats, Sprague-Dawley Sodium Chloride Comparative Study Biochemistry Molecular Biology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Cell Biology Biochemistry Molecular Biology |
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