RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development
According to previous studies, during <i>Drosophila</i> embryogenesis, the recruitment of RNA polymerase II precedes active gene transcription. This work is aimed at exploring whether this mechanism is used during <i>Drosophila</i> metamorphosis. In addition, the composition...
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MDPI AG
2022-09-01
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| Series: | International Journal of Molecular Sciences |
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| author | Marina Yu. Mazina Elena V. Kovalenko Aleksandra A. Evdokimova Maksim Erokhin Darya Chetverina Nadezhda E. Vorobyeva |
| author_facet | Marina Yu. Mazina Elena V. Kovalenko Aleksandra A. Evdokimova Maksim Erokhin Darya Chetverina Nadezhda E. Vorobyeva |
| author_sort | Marina Yu. Mazina |
| collection | DOAJ |
| description | According to previous studies, during <i>Drosophila</i> embryogenesis, the recruitment of RNA polymerase II precedes active gene transcription. This work is aimed at exploring whether this mechanism is used during <i>Drosophila</i> metamorphosis. In addition, the composition of the RNA polymerase II “paused” complexes associated with promoters at different developmental stages are described in detail. For this purpose, we performed ChIP-Seq analysis using antibodies for various modifications of RNA polymerase II (total, Pol II CTD Ser5P, and Pol II CTD Ser2P) as well as for subunits of the NELF, DSIF, and PAF complexes and Brd4/Fs(1)h that control transcription elongation. We found that during metamorphosis, similar to mid-embryogenesis, the promoters were bound by RNA polymerase II in the “paused” state, preparing for activation at later stages of development. During mid-embryogenesis, RNA polymerase II in a “pause” state was phosphorylated at Ser5 and Ser2 of Pol II CTD and bound the NELF, DSIF, and PAF complexes, but not Brd4/Fs(1)h. During metamorphosis, the “paused” RNA polymerase II complex included Brd4/Fs(1)h in addition to NELF, DSIF, and PAF. The RNA polymerase II in this complex was phosphorylated at Ser5 of Pol II CTD, but not at Ser2. These results indicate that, during mid-embryogenesis, RNA polymerase II stalls in the “post-pause” state, being phosphorylated at Ser2 of Pol II CTD (after the stage of p-TEFb action). During metamorphosis, the “pause” mechanism is closer to classical promoter-proximal pausing and is characterized by a low level of Pol II CTD Ser2P. |
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| language | English |
| last_indexed | 2024-03-09T23:45:13Z |
| publishDate | 2022-09-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-d71fb6e094394d488554d536ee6652442023-11-23T16:45:53ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-09-0123181066210.3390/ijms231810662RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> DevelopmentMarina Yu. Mazina0Elena V. Kovalenko1Aleksandra A. Evdokimova2Maksim Erokhin3Darya Chetverina4Nadezhda E. Vorobyeva5Institute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, RussiaInstitute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, RussiaInstitute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, RussiaInstitute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, RussiaInstitute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, RussiaInstitute of Gene Biology, Russian Academy of Sciences, 119334 Moscow, RussiaAccording to previous studies, during <i>Drosophila</i> embryogenesis, the recruitment of RNA polymerase II precedes active gene transcription. This work is aimed at exploring whether this mechanism is used during <i>Drosophila</i> metamorphosis. In addition, the composition of the RNA polymerase II “paused” complexes associated with promoters at different developmental stages are described in detail. For this purpose, we performed ChIP-Seq analysis using antibodies for various modifications of RNA polymerase II (total, Pol II CTD Ser5P, and Pol II CTD Ser2P) as well as for subunits of the NELF, DSIF, and PAF complexes and Brd4/Fs(1)h that control transcription elongation. We found that during metamorphosis, similar to mid-embryogenesis, the promoters were bound by RNA polymerase II in the “paused” state, preparing for activation at later stages of development. During mid-embryogenesis, RNA polymerase II in a “pause” state was phosphorylated at Ser5 and Ser2 of Pol II CTD and bound the NELF, DSIF, and PAF complexes, but not Brd4/Fs(1)h. During metamorphosis, the “paused” RNA polymerase II complex included Brd4/Fs(1)h in addition to NELF, DSIF, and PAF. The RNA polymerase II in this complex was phosphorylated at Ser5 of Pol II CTD, but not at Ser2. These results indicate that, during mid-embryogenesis, RNA polymerase II stalls in the “post-pause” state, being phosphorylated at Ser2 of Pol II CTD (after the stage of p-TEFb action). During metamorphosis, the “pause” mechanism is closer to classical promoter-proximal pausing and is characterized by a low level of Pol II CTD Ser2P.https://www.mdpi.com/1422-0067/23/18/10662transcriptionRNA polymerase IIPol II pausingPol II stallingpoised Pol II<i>Drosophila</i> |
| spellingShingle | Marina Yu. Mazina Elena V. Kovalenko Aleksandra A. Evdokimova Maksim Erokhin Darya Chetverina Nadezhda E. Vorobyeva RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development International Journal of Molecular Sciences transcription RNA polymerase II Pol II pausing Pol II stalling poised Pol II <i>Drosophila</i> |
| title | RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development |
| title_full | RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development |
| title_fullStr | RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development |
| title_full_unstemmed | RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development |
| title_short | RNA Polymerase II “Pause” Prepares Promoters for Upcoming Transcription during <i>Drosophila</i> Development |
| title_sort | rna polymerase ii pause prepares promoters for upcoming transcription during i drosophila i development |
| topic | transcription RNA polymerase II Pol II pausing Pol II stalling poised Pol II <i>Drosophila</i> |
| url | https://www.mdpi.com/1422-0067/23/18/10662 |
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