Cyclin-dependent kinase 8 module expression profiling reveals requirement of mediator subunits 12 and 13 for transcription of Serpent-dependent innate immunity genes in Drosophila

Emilia Kuuluvainen; Heini Hakala; Essi Havula; Michelle Sahal Estimé; Mika Rämet; Ville Hietakangas; Tomi P Mäkelä

doi:10.1074/jbc.M113.541904

Cyclin-dependent kinase 8 module expression profiling reveals requirement of mediator subunits 12 and 13 for transcription of Serpent-dependent innate immunity genes in Drosophila

J Biol Chem. 2014 Jun 6;289(23):16252-61. doi: 10.1074/jbc.M113.541904. Epub 2014 Apr 28.

Authors

Emilia Kuuluvainen¹, Heini Hakala¹, Essi Havula², Michelle Sahal Estimé¹, Mika Rämet³, Ville Hietakangas², Tomi P Mäkelä⁴

Affiliations

¹ From the Institute of Biotechnology, University of Helsinki, P. O. Box 56, 00014 Helsinki.
² From the Institute of Biotechnology, University of Helsinki, P. O. Box 56, 00014 Helsinki, the Department of Biosciences, University of Helsinki, P. O. Box 65, 00014 Helsinki.
³ the Institute of Biomedical Technology, and BioMediTech, University of Tampere, 33014 Tampere, the Department of Pediatrics, Tampere University Hospital, 22521 Tampere, the Department of Pediatrics, Institute of Clinical Medicine, and Medical Research Center Oulu, University of Oulu, 90014 Oulu, and the Department of Children and Adolescents, Oulu University Hospital, 90029 Oulu, Finland.
⁴ From the Institute of Biotechnology, University of Helsinki, P. O. Box 56, 00014 Helsinki, tomi.makela@helsinki.fi.

Abstract

The Cdk8 (cyclin-dependent kinase 8) module of Mediator integrates regulatory cues from transcription factors to RNA polymerase II. It consists of four subunits where Med12 and Med13 link Cdk8 and cyclin C (CycC) to core Mediator. Here we have investigated the contributions of the Cdk8 module subunits to transcriptional regulation using RNA interference in Drosophila cells. Genome-wide expression profiling demonstrated separation of Cdk8-CycC and Med12-Med13 profiles. However, transcriptional regulation by Cdk8-CycC was dependent on Med12-Med13. This observation also revealed that Cdk8-CycC and Med12-Med13 often have opposite transcriptional effects. Interestingly, Med12 and Med13 profiles overlapped significantly with that of the GATA factor Serpent. Accordingly, mutational analyses indicated that GATA sites are required for Med12-Med13 regulation of Serpent-dependent genes. Med12 and Med13 were also found to be required for Serpent-activated innate immunity genes in defense to bacterial infection. The results reveal a novel role for the Cdk8 module in Serpent-dependent transcription and innate immunity.

Keywords: Cdk8; Cyclin C; Drosophila; GATA Transcription Factor; Innate Immunity; Med12; Med13; Microarray; Serpent; Transcription.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cyclin-Dependent Kinase 8 / genetics*
Drosophila
Gene Expression Profiling*
Immunity, Innate / genetics*
Polymerase Chain Reaction
RNA Interference
Transcription, Genetic*

Substances

Cyclin-Dependent Kinase 8