An RNAi screen identifies additional members of the Drosophila Integrator complex and a requirement for cyclin C/Cdk8 in snRNA 3′-end formation

Jiandong Chen, Nader Ezzeddine, Bernhard Waltenspiel, Todd R. Albrecht, William D. Warren, William F. Marzluff, Eric Wagner

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Formation of the 3′ end of RNA polymerase II-transcribed snRNAs requires a poorly understood group of proteins called the Integrator complex. Here we used a fluorescence-based read-through reporter that expresses GFP in response to snRNA misprocessing and performed a genome-wide RNAi screen in Drosophila S2 cells to identify novel factors required for snRNA 3′-end formation. In addition to the known Integrator complex members, we identified Asunder and CG4785 as additional Integrator subunits. Functional and biochemical experiments revealed that Asunder and CG4785 are additional core members of the Integrator complex. We also identified a conserved requirement in both fly and human snRNA 3′-end processing for cyclin C and Cdk8 that is distinct from their function in the Mediator Cdk8 module. Moreover, we observed biochemical association between Integrator proteins and cyclin C/Cdk8, and that overexpression of a kinase-dead Cdk8 causes snRNA misprocessing. These data functionally define the Drosophila Integrator complex and demonstrate an additional function for cyclin C/Cdk8 unrelated to its function in Mediator.

Original languageEnglish (US)
Pages (from-to)2148-2156
Number of pages9
JournalRNA
Volume18
Issue number12
DOIs
StatePublished - Dec 2012
Externally publishedYes

Fingerprint

Cyclin C
Small Nuclear RNA
RNA Interference
Drosophila
RNA Polymerase II
Protein C
Diptera
Phosphotransferases
Fluorescence
Genome
Proteins

Keywords

  • Asunder
  • Cdk8
  • Cyclin C
  • Integrator complex
  • RNAPII
  • snRNA

ASJC Scopus subject areas

  • Molecular Biology

Cite this

An RNAi screen identifies additional members of the Drosophila Integrator complex and a requirement for cyclin C/Cdk8 in snRNA 3′-end formation. / Chen, Jiandong; Ezzeddine, Nader; Waltenspiel, Bernhard; Albrecht, Todd R.; Warren, William D.; Marzluff, William F.; Wagner, Eric.

In: RNA, Vol. 18, No. 12, 12.2012, p. 2148-2156.

Research output: Contribution to journalArticle

Chen, Jiandong ; Ezzeddine, Nader ; Waltenspiel, Bernhard ; Albrecht, Todd R. ; Warren, William D. ; Marzluff, William F. ; Wagner, Eric. / An RNAi screen identifies additional members of the Drosophila Integrator complex and a requirement for cyclin C/Cdk8 in snRNA 3′-end formation. In: RNA. 2012 ; Vol. 18, No. 12. pp. 2148-2156.
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