How do transcriptional kinases regulate cell cycle progression?

About the Project

Project:

Cyclin-dependent kinases (CDKs) are protein kinases performing essential functions by regulating cell cycle and RNA polymerase II transcription of protein-coding and non-coding genes. Each CDK requires an interaction with a specific cyclin partner for switching on their kinase activity. Mutations and dysregulation of different CDKs and associated cyclins have been linked to multiple cancers. Transcriptional CDKs, which regroups CDK7, CDK8, CDK9, CDK10, CDK11, CDK12, CDK13, and CDK19, are known to regulate gene expression via phosphorylation of RNA polymerase II and multiple associated transcription factors. Interestingly, inhibition of these transcriptional CDKs usually also affects cell cycle progression, indicating that these kinases could perform functions beyond regulation of gene expression. Understanding how and why cell cycle is affected by transcriptional CDKs inhibition will provide important insights into the roles of these kinases in cancer leading to the development of new combination therapy approaches.

In this project, the PhD student will be supervised by two labs, Dr Michael Tellier (gene expression, -omics techniques) and Dr Robert Mahen (cell cycle, microscopy). We are in possession of a series of HEK293 cell lines that have been edited with CRISPR/Cas9 to generate analogue sensitive version of multiple CDKs, including CDK7, CDK8, CDK9, CDK10, CDK12, and CDK13, allowing us to specifically and rapidly inhibit the CDK of interest with an ATP analogue. Using these cell lines, the PhD student will first investigate using flow cytometry and microscopy which transcriptional CDK is specifically regulating cell cycle progression. Based on these results, the candidate will then determine the mechanisms by which transcriptional CDKs of interest regulate cell cycle by using a combination of transcriptomics, microscopy, proteomics, and molecular and cell biology techniques. This project will therefore provide unique insights into how transcriptional CDKs can regulate cell cycle progression independently of their roles in the regulation of gene expression, providing opportunities to develop new combination therapy approaches for treating cancer where these kinases are mutated or dysregulated.

Training opportunities:

Our two labs will provide opportunities to be trained in different techniques, including human cell culture, CRISPR/Cas9, phospho-proteomics, flow cytometry, immunofluorescence, RNA-seq, nascent transcription techniques (POINT-seq, PRO-seq), bioinformatics, and standard molecular biology techniques (western blot, co-immunoprecipitation, cloning, PCR, qPCR, …). The PhD student will be able to participate in the Leicester Institute of Structural and Chemical Biology (LISCB) doctoral training programme that offer both technical and transferable career-development skills. In addition, the PhD student will have opportunities to attend conferences in UK and Europe to present his/her work. Our lab is also actively collaborating with research groups in UK, France, and Japan offering opportunities to participate in international collaborations.

Outputs:

Major outputs from the PhD project will be publications of high impact research articles that will help the applicant to be competitive in his/her future career. In addition, the applicant will be trained in multiple cutting-edge techniques (microscopy, transcriptomics, genomics, bioinformatics).

Enquiries

Project Enquiries to mt477@leicester.ac.uk

To apply please refer to

https://le.ac.uk/study/research-degrees/research-subjects/molecular-and-cell-biology