The role of Dicer protein partners in the processing of microRNA precursors.
One of the cellular functions of the ribonuclease Dicer is to process microRNA precursors (pre-miRNAs) into mature microRNAs (miRNAs). Human Dicer performs this function in cooperation with its protein partners, AGO2, PACT and TRBP. The exact role of these accessory proteins in Dicer activity is sti...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2011-01-01
|
| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3232248?pdf=render |
| _version_ | 1818512998143098880 |
|---|---|
| author | Edyta Koscianska Julia Starega-Roslan Wlodzimierz J Krzyzosiak |
| author_facet | Edyta Koscianska Julia Starega-Roslan Wlodzimierz J Krzyzosiak |
| author_sort | Edyta Koscianska |
| collection | DOAJ |
| description | One of the cellular functions of the ribonuclease Dicer is to process microRNA precursors (pre-miRNAs) into mature microRNAs (miRNAs). Human Dicer performs this function in cooperation with its protein partners, AGO2, PACT and TRBP. The exact role of these accessory proteins in Dicer activity is still poorly understood. In this study, we used the northern blotting technique to investigate pre-miRNA cleavage efficiency and specificity after depletion of AGO2, PACT and TRBP by RNAi. The results showed that the inhibition of either Dicer protein partner substantially affected not only miRNA levels but also pre-miRNA levels, and it had a rather minor effect on the specificity of Dicer cleavage. The analysis of the Dicer cleavage products generated in vitro revealed the presence of a cleavage intermediate when pre-miRNA was processed by recombinant Dicer alone. This intermediate was not observed during pre-miRNA cleavage by endogenous Dicer. We demonstrate that AGO2, PACT and TRBP were required for the efficient functioning of Dicer in cells, and we suggest that one of the roles of these proteins is to assure better synchronization of cleavages triggered by two RNase III domains of Dicer. |
| first_indexed | 2024-12-10T23:54:52Z |
| format | Article |
| id | doaj.art-486873887e894dc59109f9ae253403ff |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-10T23:54:52Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-486873887e894dc59109f9ae253403ff2022-12-22T01:28:37ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-01612e2854810.1371/journal.pone.0028548The role of Dicer protein partners in the processing of microRNA precursors.Edyta KoscianskaJulia Starega-RoslanWlodzimierz J KrzyzosiakOne of the cellular functions of the ribonuclease Dicer is to process microRNA precursors (pre-miRNAs) into mature microRNAs (miRNAs). Human Dicer performs this function in cooperation with its protein partners, AGO2, PACT and TRBP. The exact role of these accessory proteins in Dicer activity is still poorly understood. In this study, we used the northern blotting technique to investigate pre-miRNA cleavage efficiency and specificity after depletion of AGO2, PACT and TRBP by RNAi. The results showed that the inhibition of either Dicer protein partner substantially affected not only miRNA levels but also pre-miRNA levels, and it had a rather minor effect on the specificity of Dicer cleavage. The analysis of the Dicer cleavage products generated in vitro revealed the presence of a cleavage intermediate when pre-miRNA was processed by recombinant Dicer alone. This intermediate was not observed during pre-miRNA cleavage by endogenous Dicer. We demonstrate that AGO2, PACT and TRBP were required for the efficient functioning of Dicer in cells, and we suggest that one of the roles of these proteins is to assure better synchronization of cleavages triggered by two RNase III domains of Dicer.http://europepmc.org/articles/PMC3232248?pdf=render |
| spellingShingle | Edyta Koscianska Julia Starega-Roslan Wlodzimierz J Krzyzosiak The role of Dicer protein partners in the processing of microRNA precursors. PLoS ONE |
| title | The role of Dicer protein partners in the processing of microRNA precursors. |
| title_full | The role of Dicer protein partners in the processing of microRNA precursors. |
| title_fullStr | The role of Dicer protein partners in the processing of microRNA precursors. |
| title_full_unstemmed | The role of Dicer protein partners in the processing of microRNA precursors. |
| title_short | The role of Dicer protein partners in the processing of microRNA precursors. |
| title_sort | role of dicer protein partners in the processing of microrna precursors |
| url | http://europepmc.org/articles/PMC3232248?pdf=render |
| work_keys_str_mv | AT edytakoscianska theroleofdicerproteinpartnersintheprocessingofmicrornaprecursors AT juliastaregaroslan theroleofdicerproteinpartnersintheprocessingofmicrornaprecursors AT wlodzimierzjkrzyzosiak theroleofdicerproteinpartnersintheprocessingofmicrornaprecursors AT edytakoscianska roleofdicerproteinpartnersintheprocessingofmicrornaprecursors AT juliastaregaroslan roleofdicerproteinpartnersintheprocessingofmicrornaprecursors AT wlodzimierzjkrzyzosiak roleofdicerproteinpartnersintheprocessingofmicrornaprecursors |