Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs

Victoria A. Church, Sigal Pressman, Mamiko Isaji, Mary Truscott, Nihal Terzi Cizmecioglu, Stephen Buratowski, Maxim V. Frolov, Richard W Carthew*

*Corresponding author for this work

Research output: Contribution to journalArticle

7 Citations (Scopus)

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 languageEnglish (US)
Pages (from-to)3123-3134
Number of pages12
JournalCell reports
Volume20
Issue number13
DOIs
StatePublished - Sep 26 2017

Fingerprint

RNA Polymerase II
Microcomputers
MicroRNAs
Microprocessor chips
Processing
Association reactions
Phosphorylation
DNA-Directed RNA Polymerases
Transcription
Drosophila
Elongation
Phosphotransferases
RNA

Keywords

  • DGCR8
  • Drosophila
  • RNA polymerase II
  • microRNA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Church, Victoria A. ; Pressman, Sigal ; Isaji, Mamiko ; Truscott, Mary ; Cizmecioglu, Nihal Terzi ; Buratowski, Stephen ; Frolov, Maxim V. ; Carthew, Richard W. / Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs. In: Cell reports. 2017 ; Vol. 20, No. 13. pp. 3123-3134.
@article{453ac36e6ffa4023a15dc09143c61773,
title = "Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs",
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.",
keywords = "DGCR8, Drosophila, RNA polymerase II, microRNA",
author = "Church, {Victoria A.} and Sigal Pressman and Mamiko Isaji and Mary Truscott and Cizmecioglu, {Nihal Terzi} and Stephen Buratowski and Frolov, {Maxim V.} and Carthew, {Richard W}",
year = "2017",
month = "9",
day = "26",
doi = "10.1016/j.celrep.2017.09.010",
language = "English (US)",
volume = "20",
pages = "3123--3134",
journal = "Cell Reports",
issn = "2211-1247",
publisher = "Cell Press",
number = "13",

}

Church, VA, Pressman, S, Isaji, M, Truscott, M, Cizmecioglu, NT, Buratowski, S, Frolov, MV & Carthew, RW 2017, 'Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs', Cell reports, vol. 20, no. 13, pp. 3123-3134. https://doi.org/10.1016/j.celrep.2017.09.010

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 journalArticle

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 -

Church VA, Pressman S, Isaji M, Truscott M, Cizmecioglu NT, Buratowski S et al. Microprocessor Recruitment to Elongating RNA Polymerase II Is Required for Differential Expression of MicroRNAs. Cell reports. 2017 Sep 26;20(13):3123-3134. https://doi.org/10.1016/j.celrep.2017.09.010