RNA polymerase II (RNAPII) directs transcription of protein coding genes and this process consists of several stages including preinitiation complex formation, productive elongation and termination. This transcription cycle is tightly linked to co-transcriptional maturation of nascent transcripts including pre-mRNA splicing and polyadenylation. RNAPII contains an unstructured C-terminal domain (CTD) with repeats of evolutionarily conserved heptapeptide YSPTSPS, where individual serines get phosphorylated. Several cyclin-dependent kinases (Cdks) regulate the phosphorylation status of the CTD and subsequent binding of transcription and pre-mRNA processing factors. Thus, the patterns of phosphorylation of the CTD direct actions of RNAPII during transcriptional cycle and co-transcriptional processing of nascent transcripts. Moreover, CTD was also functionally linked to DNA damage response and maintenance of genome stability via regulation of transcription, mRNA processing and recombination. Thus, CTD and its posttranslational modifications, associated proteins and modifying enzymes are emerging as new players in cellular response to DNA damage. Our recent work led to the identification of the Cyclin K/Cdk12 complex that phosphorylates serine 2 in the CTD of RNAPII and directs expression of several crucial DNA damage response genes including BRCA1, ATR or FANCI. In my lab we apply a combination of biochemical, proteomics and genome-wide techniques to determine the molecular mechanism that regulates the expression of Cdk12-dependent genes with a focus on DNA damage response genes. The ultimate goal of our research is to uncover how the CycK/Cdk12 complex and the CTD of RNAPII contribute to the maintenance of genome stability, and how disruption of their functions lead to the onset of a malignant state.
list / cards
Name and position |
Phone |
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Dalibor Blažek, Ph.D. Research Group Leader |
+420 54949 6649, +420 54949 7564 | |
Pavla Gajdušková, Ph.D. Researcher |
+420 54949 7566 | |
Květa Pilařová PhD student |
+420 54949 7566 | |
Anil Paul Chirackal Manavalan, Ph.D. odborný pracovník ve výzkumu - postdoc |
+420 54949 7566 | |
Michal Rájecký, Ph.D. PhD student |
+420 54949 6649, +420 54949 7564 | |
Martin Bednařík Specialist |
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Přemysl Souček, Ph.D. senior researcher |
+420 54949 7567 | |
Milan Hluchý PhD student |
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Jan Herudek odborný pracovník ve výzkumu - postdoc |
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Lenka Humpolíková Adámková, Ph.D. odborná pracovnice |
+420 54949 3878 | |
Zuzana Slabá odborná pracovnice |
+420 54949 4443 | |
Kateřina Hanáková Research specialist - PhD student |
+420 54949 8442 |
Shared laboratory: A35/111, phone: 549 49 6649
Laboratories equipped with technologies used in molecular biological analysis: RealTime PCR cyclers, PCR cyclers, laminar boxes, CO2 boxes.
Supervisor: Mgr. Dalibor Blažek, Ph.D.
Consultant: doc. RNDr. Zbyněk Zdráhal, Dr.
Cdk12 and Cdk13 are transcriptional cyclin-dependent kinases (Cdk) found mutated in various cancers. In previous studies we found that Cdk12 maintains genome stability via optimal transcription of key homologous recombination repair pathway genes including BRCA1. Cellular function of Cdk13 remains unclear. Apart from the C-terminal domain of RNA Polymerase II other cellular substrates for both kinases are not known. In this research we propose using a screen in cells carrying an analog sensitive mutants of both kinases to discover their novel cellular substrates. The substrates and their roles in normal and cancerous cells will be characterized by various techniques of molecular biology and biochemistry.
Keywords: cyclin-dependent kinases, Cdk12, Cdk13, recombination repair pathway genes, BRCA1, cancer
Supervisor: Mgr. Dalibor Blažek, Ph.D.
Consultant: doc. RNDr. Zbyněk Zdráhal, Dr.
Gene expression of protein-coding genes is regulated at many levels. This project focuses on transcription cycle-related cyclin-dependent kinase 12 (Cdk12). We showed Cdk12 to be involved in the regulation of gene expression of key DNA damage response genes (BRCA1, FANCI, ATR) via phosphorylation of the C-terminal domain of RNA Polymerase II (see our recent papers: Blazek et al., Genes and Development 2011; 25(20):2158-72; Bosken et al. Nature Comm. 2014;24;5:3505; Ekumi at al., Nucleic Acids Research 2015;11;43(5):2575-89). Also, Cdk12 was found among the most often mutated genes in ovarian carcinoma (Ovarian Carcinoma Cancer Genome Atlas, Nature, 2011). In accordance with the proposed broad role of the Cdk12 in the regulation of gene expression/DNA damage response is its deregulation in many other cancers. However, mechanism how Cdk12 contributes to the regulation of gene expression is unknown. The main aim of the project will be deciphering of molecular mechanism that mediates expression of Cdk12-dependent genes. To address this question many techniques of molecular biology and biochemistry will be applied including state of the art mass spectrometry and next generation sequencing (ChIP-seq etc.).Ovarian carcinoma cell lines will be used as a model for the studies.
Keywords: cyclin-dependent kinase 12, Cdk12, regulation of gene expression, ovarian carcinoma, DNA damage response, mass spektrometry, next generation sequencing
29. ledna 2018 9:46
LECTURE: Dr. Ondrej Hovorka: Models of magnetic nanoparticles for biomedical applications
25. ledna 2018 18:21
WHEN: 30. 01. 2018 WHERE: CEITEC BUT, Purkynova 123, large meeting room SPEAKER: Dr Andriy Marko TALK: Advances in PELDOR…