Advancing Epigenetics Towards Systems Biology

Building a high-resolution map of protein interactions along the RNA polymerase II CTD using site-specific BPA crosslinking

IRCM

Description

Project summary
RNA polymerase II, the enzyme responsible for the transcription of pre-mRNA and most non-coding RNAs in eukaryotes, contains a carboxy-terminal domain (CTD) made of repetitions of a heptapeptide having YSPTSPS as consensus sequence. This peptide is repeated 26 times in yeast and 52 times in human. The tyrosine (Tyr1), threonine (Thr4) and all three serines (Ser2, 5 and 7) of each repetition are subject to a cycle of phosphorylation and dephosphorylation during transcription, creating an astronomical number of possible combinatorial phosphorylation states that are thought to constitute a “CTD code” read by CTD-binding proteins. Long-standing questions in the field are whether individual repeats have specific functions and whether the “CTD proteome” assembles in an organized structure or consists of very dynamic low affinity and high valency interactions? In this project, we propose to use BPA crosslinking to create a high-resolution map of protein binding along the CTD in living cells as a way to molecularly decipher the functions of different CTD repeats or regions.

Summary of responsibilities
This project takes advantage of clever manipulations of the genetic code in Saccharomyces cerevisiae to introduce a modified amino acid (BPA) that can be induced to crosslink neighboring proteins in specific regions of the CTD in living cells. This approach will be used in combination with state-of-the-art proteomic technologies and standard biochemistry and molecular biology methods. The postdoc will be in charge of executing and analysing the vast majority of the experiments for the project and will be assisted during his/her training by Dr. Robert and senior members of the lab. The IRCM has several core laboratories with expertise that will enhance the training experience of the candidate.

Recent selected publications

  • Collin P, Jeronimo C, Poitras C, Robert F. (2018) RNA Polymerase II CTD Tyrosine 1 is Required for Efficient Termination by the Nrd1-Nab3-Sen1 Pathway. Under review.
  • Jeronimo C, Collin P, Robert F. (2016) The RNA Polymerase II CTD: The Increasing Complexity of a Low-Complexity Protein Domain. J Mol Biol. 2016 Jun 19;428(12):2607-2622.
  • Bataille AR, Jeronimo C, Jacques PÉ, Laramée L, Fortin MÈ, Forest A, Bergeron M, Hanes SD, Robert F. (2012) A universal RNA polymerase II CTD cycle is orchestrated by complex interplays between kinase, phosphatase, and isomerase enzymes along genes. Mol Cell. 2012 Jan 27;45(2):158-70.

Qualifications

We seek a highly motivated individual with a genuine interest in understanding the mechanistic aspects of molecular processes. A background in molecular biology and/or biochemistry is mandatory. Knowledge in bioinformatics, computer programming or statistics are assets but not mandatory. Only candidates with very good academic track records will be considered.

Salary

40,000$ per year

Application details

Please apply by sending your CV and publication list together with a motivation letter to:
Francois Robert - This email address is being protected from spambots. You need JavaScript enabled to view it.

Relevant web link: https://www.robertlab.org

Starting date: 31st October 2018
Deadline for applications: 1st April 2019