Exonuclease resistant 18S and 25S ribosomal RNA components in yeast are possibly newly transcribed by RNA polymerase II
| Autor: | Miguel A. Rocha, Jacob Fleischmann, Peter Hauser, Bhavani S. Gowda, Mary Grace D. Pilapil |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Exonucleases
Exonuclease Transcription Genetic RNA polymerase II DNA Ribosomal Primer extension Fungal Proteins 03 medical and health sciences 0302 clinical medicine Candida albicans RNA Ribosomal 18S RNA polymerase I lcsh:QH573-671 Molecular Biology Polymerase 030304 developmental biology 0303 health sciences Base Sequence biology Chemistry lcsh:Cytology RNA RNA Fungal Cell Biology Ribosomal RNA Biochemistry RNA Ribosomal 030220 oncology & carcinogenesis biology.protein RNA Polymerase II Transcription factor II B Research Article |
| Zdroj: | BMC Molecular and Cell Biology, Vol 21, Iss 1, Pp 1-12 (2020) BMC Molecular and Cell Biology |
| ISSN: | 2661-8850 |
| DOI: | 10.1186/s12860-020-00303-z |
| Popis: | Background We have previously reported 18S and 25S ribosomal RNA molecules in Candida albicans resistant to processive 5′ → 3′ exonuclease, appearing as cells approached stationary growth phase. Initial analysis pointed to extra phosphate(s) at their 5′- end raising the possibility that they were newly transcribed. Here we report on additional experiments exploring this possibility and try to establish which of the RNA polymerases may be transcribing them. Results Oligo-ligation and primer extension again showed the presence of extra phosphate at the 5′-end of the reported processing sites for both 18S and 25S ribosomal RNA components. Inhibition of Pol I with BMH-21 increased the presence of the molecules. Quantitation with an Agilent Bioanalyzer showed that resistant 18S and 25S molecules are primarily produced in the nucleus. Utilizing an RNA cap specific antibody, a signal could be detected on these molecules via immunoblotting; such signal could be eliminated by decapping reaction. Both the cap specific antibody and eIF4E cap-binding protein, increased fold enrichment upon quantitative amplification. Antibodies specific for the RNA Polymerase II c-terminal domain and TFIIB initiator factor showed the presence of Pol II on DNA sequences for both 18S and 25S molecules in chromatin precipitation and qPCR assays. Rapamycin inhibition of TOR complex also resulted in an increase of resistant 18S and 25S molecules. Conclusions These data raise the possibility of a role for RNA Polymerase II in the production of 18S and 25S molecules and indicate that efforts for more direct proof may be worthwhile. If definitively proven it will establish an additional role for RNA Polymerase II in ribosomal production. |
| Databáze: | OpenAIRE |
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