10779/crick.11371614.v1
Kerstin Köhler
Kerstin
Köhler
Luis Sanchez-Pulido
Luis
Sanchez-Pulido
Verena Höfer
Verena
Höfer
Anika Marko
Anika
Marko
Chris P Ponting
Chris P
Ponting
Ambrosius P Snijders
Ambrosius P
Snijders
Regina Feederle
Regina
Feederle
Aloys Schepers
Aloys
Schepers
Dominik Boos
Dominik
Boos
The Cdk8/19-cyclin C transcription regulator functions in genome replication through metazoan Sld7
The Francis Crick Institute
2019
Carrier Proteins
Cell Cycle Proteins
Computational Biology
Cyclin C
Cyclin-Dependent Kinase 8
Cyclin-Dependent Kinases
DNA Replication
DNA-Binding Proteins
HEK293 Cells
HeLa Cells
Humans
Mitosis
Protein Binding
Saccharomyces cerevisiae
Saccharomyces cerevisiae Proteins
Hela Cells
PRT
06 Biological Sciences
11 Medical and Health Sciences
07 Agricultural and Veterinary Sciences
Developmental Biology
2019-12-16 17:40:36
Journal contribution
https://crick.figshare.com/articles/journal_contribution/The_Cdk8_19-cyclin_C_transcription_regulator_functions_in_genome_replication_through_metazoan_Sld7/11371614
Accurate genome duplication underlies genetic homeostasis. Metazoan Mdm2 binding protein (MTBP) forms a main regulatory platform for origin firing together with Treslin/TICRR and TopBP1 (Topoisomerase II binding protein 1 (TopBP1)-interacting replication stimulating protein/TopBP1-interacting checkpoint and replication regulator). We report the first comprehensive analysis of MTBP and reveal conserved and metazoa-specific MTBP functions in replication. This suggests that metazoa have evolved specific molecular mechanisms to adapt replication principles conserved with yeast to the specific requirements of the more complex metazoan cells. We uncover one such metazoa-specific process: a new replication factor, cyclin-dependent kinase 8/19-cyclinC (Cdk8/19-cyclin C), binds to a central domain of MTBP. This interaction is required for complete genome duplication in human cells. In the absence of MTBP binding to Cdk8/19-cyclin C, cells enter mitosis with incompletely duplicated chromosomes, and subsequent chromosome segregation occurs inaccurately. Using remote homology searches, we identified MTBP as the metazoan orthologue of yeast synthetic lethal with Dpb11 7 (Sld7). This homology finally demonstrates that the set of yeast core factors sufficient for replication initiation in vitro is conserved in metazoa. MTBP and Sld7 contain two homologous domains that are present in no other protein, one each in the N and C termini. In MTBP the conserved termini flank the metazoa-specific Cdk8/19-cyclin C binding region and are required for normal origin firing in human cells. The N termini of MTBP and Sld7 share an essential origin firing function, the interaction with Treslin/TICRR or its yeast orthologue Sld3, respectively. The C termini may function as homodimerisation domains. Our characterisation of broadly conserved and metazoa-specific initiation processes sets the basis for further mechanistic dissection of replication initiation in vertebrates. It is a first step in understanding the distinctions of origin firing in higher eukaryotes.