Vesela Encheva,2 Ilaria Gori,3 Rebecca E. Saunders,3 Rachael Instrell,3 Ozan Aygun,1,7 Marta Rodriguez-Martinez,1 Juston C. Weems,4 Gavin P. Kelly,5 Joan W. Conaway,4,6 Ronald C. Conaway,4,6 Aengus Stewart,5 Michael Howell,3 Ambrosius P. Snijders,2 and Jesper Q. Svejstrup1,*of Transcription Laboratory, the Francis Crick Institute, Clare Hall Laboratories, South Mimms EN6 3LD, UK Analysis and Proteomics Laboratory, the Francis Crick Institute, Clare Hall Laboratories, South Mimms EN6 3LD, UK 3High Throughput Screening Laboratory, the Francis Crick Institute, 44 Lincoln’s Inn Imatinib (Mesylate) price Fields, London WC2A 3LY, UK 4Stowers Grazoprevir site Institute for Medical Research, Kansas City, MO 64110, USA 5Bioinformatics and Biostatistics Laboratory, the Francis Crick Institute, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK 6Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA 7Present address: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA *Correspondence: [email protected] http://dx.doi.org/10.1016/j.celrep.2016.04.2Protein 1MechanismsSUMMARYIn order to facilitate the identification of factors and pathways in the cellular response to UV-induced DNA damage, several descriptive proteomic screens and a functional genomics screen were performed in parallel. Numerous factors could be identified with high confidence when the screen results were superimposed and interpreted together, incorporating biological knowledge. A searchable database, bioLOGIC, which provides access to relevant information about a protein or process of interest, was established to host the results and facilitate data mining. Besides uncovering roles in the DNA damage response for numerous proteins and complexes, including Integrator, Cohesin, PHF3, ASC-1, SCAF4, SCAF8, and SCAF11, we uncovered a role for the poorly studied, melanomaassociated serine/threonine kinase 19 (STK19). Besides effectively uncovering relevant factors, the multiomic approach also provides a systems-wide overview of the diverse cellular processes connected to the transcription-related DNA damage response.INTRODUCTION The cellular response to bulky DNA lesions, such as those induced by UV irradiation is multi-faceted. The effect of such damage on transcription is particularly complex. Bulky DNA lesions in the transcribed strand cause stalling of RNA polymerase II (RNAPII), resulting in a block to transcript elongation. Damagestalled RNAPII then functions as a molecular beacon that triggers transcription-coupled nucleotide excision repair (TC-NER), the process whereby DNA damage in the transcribed strand of active genes is preferentially removed (Gaillard and Aguilera, 2013). On the other hand, if the DNA lesion for some reason cannot be removed by TC-NER, a mechanism of last resort en-sures that RNAPII is ubiquitylated and degraded by the proteasome, enabling repair by other mechanisms (Wilson et al., 2013). Importantly, bulky DNA lesions not only block RNAPII progress, but also affect transcription genome-wide so that even un-damaged genes temporarily cease to be expressed (Mayne and Lehmann, 1982; Rockx et al., 2000; Proietti-De-Santis et al., 2006). The mechanisms and factors that underlie TCNER and the more general DNA-damage-induced repression of gene expression are still poorly understood. Cockayne syndrome B protein (CSB, also named ERCC6) plays a key role in both TC-NER and the global transcription response to DNA.Vesela Encheva,2 Ilaria Gori,3 Rebecca E. Saunders,3 Rachael Instrell,3 Ozan Aygun,1,7 Marta Rodriguez-Martinez,1 Juston C. Weems,4 Gavin P. Kelly,5 Joan W. Conaway,4,6 Ronald C. Conaway,4,6 Aengus Stewart,5 Michael Howell,3 Ambrosius P. Snijders,2 and Jesper Q. Svejstrup1,*of Transcription Laboratory, the Francis Crick Institute, Clare Hall Laboratories, South Mimms EN6 3LD, UK Analysis and Proteomics Laboratory, the Francis Crick Institute, Clare Hall Laboratories, South Mimms EN6 3LD, UK 3High Throughput Screening Laboratory, the Francis Crick Institute, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK 4Stowers Institute for Medical Research, Kansas City, MO 64110, USA 5Bioinformatics and Biostatistics Laboratory, the Francis Crick Institute, 44 Lincoln’s Inn Fields, London WC2A 3LY, UK 6Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City, KS 66160, USA 7Present address: Department of Chemistry, Massachusetts Institute of Technology, Cambridge, MA 02139, USA *Correspondence: [email protected] http://dx.doi.org/10.1016/j.celrep.2016.04.2Protein 1MechanismsSUMMARYIn order to facilitate the identification of factors and pathways in the cellular response to UV-induced DNA damage, several descriptive proteomic screens and a functional genomics screen were performed in parallel. Numerous factors could be identified with high confidence when the screen results were superimposed and interpreted together, incorporating biological knowledge. A searchable database, bioLOGIC, which provides access to relevant information about a protein or process of interest, was established to host the results and facilitate data mining. Besides uncovering roles in the DNA damage response for numerous proteins and complexes, including Integrator, Cohesin, PHF3, ASC-1, SCAF4, SCAF8, and SCAF11, we uncovered a role for the poorly studied, melanomaassociated serine/threonine kinase 19 (STK19). Besides effectively uncovering relevant factors, the multiomic approach also provides a systems-wide overview of the diverse cellular processes connected to the transcription-related DNA damage response.INTRODUCTION The cellular response to bulky DNA lesions, such as those induced by UV irradiation is multi-faceted. The effect of such damage on transcription is particularly complex. Bulky DNA lesions in the transcribed strand cause stalling of RNA polymerase II (RNAPII), resulting in a block to transcript elongation. Damagestalled RNAPII then functions as a molecular beacon that triggers transcription-coupled nucleotide excision repair (TC-NER), the process whereby DNA damage in the transcribed strand of active genes is preferentially removed (Gaillard and Aguilera, 2013). On the other hand, if the DNA lesion for some reason cannot be removed by TC-NER, a mechanism of last resort en-sures that RNAPII is ubiquitylated and degraded by the proteasome, enabling repair by other mechanisms (Wilson et al., 2013). Importantly, bulky DNA lesions not only block RNAPII progress, but also affect transcription genome-wide so that even un-damaged genes temporarily cease to be expressed (Mayne and Lehmann, 1982; Rockx et al., 2000; Proietti-De-Santis et al., 2006). The mechanisms and factors that underlie TCNER and the more general DNA-damage-induced repression of gene expression are still poorly understood. Cockayne syndrome B protein (CSB, also named ERCC6) plays a key role in both TC-NER and the global transcription response to DNA.
