Abstract
The cellular abundance of mature microRNAs (miRNAs) is dictated by the efficiency of nuclear processing of primary miRNA transcripts (pri-miRNAs) into pre-miRNA intermediates. The Microprocessor complex of Drosha and DGCR8 carries this out, but it has been unclear what controls Microprocessor's differential processing of various pri-miRNAs. Here, we show that Drosophila DGCR8 (Pasha) directly associates with the C-terminal domain of the RNA polymerase II elongation complex when it is phosphorylated by the Cdk9 kinase (pTEFb). When association is blocked by loss of Cdk9 activity, a global change in pri-miRNA processing is detected. Processing of pri-miRNAs with a UGU sequence motif in their apical junction domain increases, while processing of pri-miRNAs lacking this motif decreases. Therefore, phosphorylation of RNA polymerase II recruits Microprocessor for co-transcriptional processing of non-UGU pri-miRNAs that would otherwise be poorly processed. In contrast, UGU-positive pri-miRNAs are robustly processed by Microprocessor independent of RNA polymerase association. Nuclear processing of microRNAs is a major determinant of cellular abundance of these RNAs. Church et al. find that the DGCR8 subunit of Microprocessor binds to RNA polymerase II. This couples microRNA processing to transcription. If microRNAs lack a sequence motif, co-transcriptional processing plays a more important role in determining abundance.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 3123-3134 |
| Number of pages | 12 |
| Journal | Cell reports |
| Volume | 20 |
| Issue number | 13 |
| DOIs | |
| State | Published - Sep 26 2017 |
Fingerprint
Keywords
- DGCR8
- Drosophila
- RNA polymerase II
- microRNA
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
}
Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs. / Church, Victoria A.; Pressman, Sigal; Isaji, Mamiko; Truscott, Mary; Cizmecioglu, Nihal Terzi; Buratowski, Stephen; Frolov, Maxim V.; Carthew, Richard W.
In: Cell reports, Vol. 20, No. 13, 26.09.2017, p. 3123-3134.Research output: Contribution to journal › Article
TY - JOUR
T1 - Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs
AU - Church, Victoria A.
AU - Pressman, Sigal
AU - Isaji, Mamiko
AU - Truscott, Mary
AU - Cizmecioglu, Nihal Terzi
AU - Buratowski, Stephen
AU - Frolov, Maxim V.
AU - Carthew, Richard W
PY - 2017/9/26
Y1 - 2017/9/26
N2 - The cellular abundance of mature microRNAs (miRNAs) is dictated by the efficiency of nuclear processing of primary miRNA transcripts (pri-miRNAs) into pre-miRNA intermediates. The Microprocessor complex of Drosha and DGCR8 carries this out, but it has been unclear what controls Microprocessor's differential processing of various pri-miRNAs. Here, we show that Drosophila DGCR8 (Pasha) directly associates with the C-terminal domain of the RNA polymerase II elongation complex when it is phosphorylated by the Cdk9 kinase (pTEFb). When association is blocked by loss of Cdk9 activity, a global change in pri-miRNA processing is detected. Processing of pri-miRNAs with a UGU sequence motif in their apical junction domain increases, while processing of pri-miRNAs lacking this motif decreases. Therefore, phosphorylation of RNA polymerase II recruits Microprocessor for co-transcriptional processing of non-UGU pri-miRNAs that would otherwise be poorly processed. In contrast, UGU-positive pri-miRNAs are robustly processed by Microprocessor independent of RNA polymerase association. Nuclear processing of microRNAs is a major determinant of cellular abundance of these RNAs. Church et al. find that the DGCR8 subunit of Microprocessor binds to RNA polymerase II. This couples microRNA processing to transcription. If microRNAs lack a sequence motif, co-transcriptional processing plays a more important role in determining abundance.
AB - The cellular abundance of mature microRNAs (miRNAs) is dictated by the efficiency of nuclear processing of primary miRNA transcripts (pri-miRNAs) into pre-miRNA intermediates. The Microprocessor complex of Drosha and DGCR8 carries this out, but it has been unclear what controls Microprocessor's differential processing of various pri-miRNAs. Here, we show that Drosophila DGCR8 (Pasha) directly associates with the C-terminal domain of the RNA polymerase II elongation complex when it is phosphorylated by the Cdk9 kinase (pTEFb). When association is blocked by loss of Cdk9 activity, a global change in pri-miRNA processing is detected. Processing of pri-miRNAs with a UGU sequence motif in their apical junction domain increases, while processing of pri-miRNAs lacking this motif decreases. Therefore, phosphorylation of RNA polymerase II recruits Microprocessor for co-transcriptional processing of non-UGU pri-miRNAs that would otherwise be poorly processed. In contrast, UGU-positive pri-miRNAs are robustly processed by Microprocessor independent of RNA polymerase association. Nuclear processing of microRNAs is a major determinant of cellular abundance of these RNAs. Church et al. find that the DGCR8 subunit of Microprocessor binds to RNA polymerase II. This couples microRNA processing to transcription. If microRNAs lack a sequence motif, co-transcriptional processing plays a more important role in determining abundance.
KW - DGCR8
KW - Drosophila
KW - RNA polymerase II
KW - microRNA
UR - http://www.scopus.com/inward/record.url?scp=85029843704&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85029843704&partnerID=8YFLogxK
U2 - 10.1016/j.celrep.2017.09.010
DO - 10.1016/j.celrep.2017.09.010
M3 - Article
C2 - 28954229
AN - SCOPUS:85029843704
VL - 20
SP - 3123
EP - 3134
JO - Cell Reports
JF - Cell Reports
SN - 2211-1247
IS - 13
ER -