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., and Wagner, Eric J. (2012) An RNAi screen identifies additional members of the Drosophila Integrator complex and a requirement for cyclin C/Cdk8 in snRNA 3'-end formation. RNA, 18 (12). pp. 2148-2156.

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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.

Item ID: 25840
Item Type: Article (Research - C1)
ISSN: 1469-9001
Keywords: integrator complex, snRNA, RNAPII, cyclin C, Cdk8, Asunder
Funders: Queensland Cancer Fund and the NHMRC through the Australian Drosophila Biomedical Research Support Facility.
Date Deposited: 22 Mar 2013 03:46
FoR Codes: 06 BIOLOGICAL SCIENCES > 0601 Biochemistry and Cell Biology > 060103 Cell Development, Proliferation and Death @ 50%
06 BIOLOGICAL SCIENCES > 0604 Genetics > 060407 Genome Structure and Regulation @ 25%
06 BIOLOGICAL SCIENCES > 0604 Genetics > 060405 Gene Expression (incl Microarray and other genome-wide approaches) @ 25%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970106 Expanding Knowledge in the Biological Sciences @ 90%
93 EDUCATION AND TRAINING > 9399 Other Education and Training > 939999 Education and Training not elsewhere classified @ 10%
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