Article ; Online: DEAD-box RNA helicase Dbp4 is required for small-subunit processome formation and function.
Molecular and cellular biology
2015 Volume 35, Issue 5, Page(s) 816–830
Abstract: DEAD-box RNA helicase Dbp4 is required for 18S rRNA synthesis: cellular depletion of Dbp4 impairs ... with the functionally active U3 RNP; this particle, called the small-subunit (SSU) processome, can be observed at the 5' end ... the early cleavage reactions of the pre-rRNA and causes U14 small nucleolar RNA (snoRNA) to remain ...
| Abstract | DEAD-box RNA helicase Dbp4 is required for 18S rRNA synthesis: cellular depletion of Dbp4 impairs the early cleavage reactions of the pre-rRNA and causes U14 small nucleolar RNA (snoRNA) to remain associated with pre-rRNA. Immunoprecipitation experiments (IPs) carried out with whole-cell extracts (WCEs) revealed that hemagglutinin (HA)-tagged Dbp4 is associated with U3 snoRNA but not with U14 snoRNA. IPs with WCEs also showed association with the U3-specific protein Mpp10, which suggests that Dbp4 interacts with the functionally active U3 RNP; this particle, called the small-subunit (SSU) processome, can be observed at the 5' end of nascent pre-rRNA. Electron microscopy analyses indicated that depletion of Dbp4 compromised SSU processome formation and cotranscriptional cleavage of the pre-rRNA. Sucrose density gradient analyses revealed that depletion of U3 snoRNA or the Mpp10 protein inhibited the release of U14 snoRNA from pre-rRNA, just as was seen with Dbp4-depleted cells, indicating that alteration of SSU processome components has significant consequences for U14 snoRNA dynamics. We also found that the C-terminal extension flanking the catalytic core of Dbp4 plays an important role in the release of U14 snoRNA from pre-rRNA. |
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| MeSH term(s) | Centrifugation, Density Gradient ; Chromatin/chemistry ; DEAD-box RNA Helicases/metabolism ; Genotype ; Microscopy, Electron ; Phosphoproteins/metabolism ; Protein Structure, Tertiary ; RNA Helicases/metabolism ; RNA Nucleotidyltransferases/metabolism ; RNA, Ribosomal, 18S/metabolism ; RNA, Small Nucleolar/metabolism ; Ribonucleoproteins/metabolism ; Ribosomes/chemistry ; Saccharomyces cerevisiae/genetics ; Saccharomyces cerevisiae Proteins/metabolism |
| Chemical Substances | Chromatin ; MPP10 protein, S cerevisiae ; Phosphoproteins ; RNA, Ribosomal, 18S ; RNA, Small Nucleolar ; RNA, U3 small nucleolar ; Ribonucleoproteins ; Saccharomyces cerevisiae Proteins ; RNA Nucleotidyltransferases (EC 2.7.7.-) ; HCA4 protein, S cerevisiae (EC 3.6.1.-) ; DEAD-box RNA Helicases (EC 3.6.4.13) ; RNA Helicases (EC 3.6.4.13) |
| Language | English |
| Publishing date | 2015-03 |
| Publishing country | United States |
| Document type | Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S. |
| ZDB-ID | 779397-2 |
| ISSN | 1098-5549 ; 0270-7306 |
| ISSN (online) | 1098-5549 |
| ISSN | 0270-7306 |
| DOI | 10.1128/MCB.01348-14 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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