PRESENTATION OF THE SUB-PROJECT 5
Role of protein secretion in adaptation of L. monocytogenes
Host organisation: INRA Secondments: University of Burgundy (UB) and University of Copenhagen (UCPH)
HOST INSTITUTION AND SUPERVISOR:
INRA UR454, DR. Michel HÉBRAUD (more details)
OBJECTIVES:
This project will connect the role of secreted protein, particularly the cell surface-associated proteins named surfaceome (or surfome) and the extracellular proteins named exoproteome during biofilm formation and adaptation to controlled environments (eg low air relative humidity). A combination of classical and innovative proteomic approaches by using different protein extraction methods (precipitation, cell fractionation, cell enzymatic shaving), in-gel and off-gel protein separation and analysis (2-DE and Maldi-Tof MS; label-free quantitative LC-MS/MS) and Maldi imaging MS of proteins, peptides and lipids in biofilms will be implemented to explore the molecular response of L. monocytogenes. The analyses will be performed both with a reference sequenced strain and a strain qualified as persistent due to its identification during several years in a foodstuff plant (but never found on output food products). Deletion mutants will be constructed to understand the regulation and role of some proteins of interest.
EXPECTED RESULTS:
Quantitative proteome allowing linkage between mRNA levels and protein levels in order to strenghten interpretation of transcriptome data. Decipher molecilar response in the interface cell/environment
SECONDMENTS:
University of Copenhagen (more details) and University of Burgundy (more details)
DURATION OF THE RECRUITMENT :
36 months
PRESENTATION OF THE FELLOW
Tiago SANTOS
MY PREVIOUS TRAINING AND EXPERIENCE:
I’m a 25 years old researcher with an h-index of 4 and more than 4 years lab experience that culminated in 8 published papers, 2 chapters and 30 communications in scientific meetings. I carried out my studies in Portugal, at the University of Trás-os-Montes and Alto Douro (UTAD), where I received, in 2012, my Bachelor of Science degree in Genetics and Biotechnology. There, I acquired my first research experience and my strong interest in molecular microbiology, and my will to prosecute my career on general proteomics. Afterwards, I decided to continue my academic training in the same institution, with a Master Degree in Molecular, Comparative and Technological Genetics. After my master’s (May 2014) I continued my collaboration in the Center of Genetics and Biotechnology (CGB-UTAD). In July 2014 I won a one-year MSc research fellow in the INNOFOOD project (CGB-UTAD). Research work I developed up to date was focused on the use of genomics and proteomics in microbial species and plant varieties.
WHY I WANTED TO JOIN THE PROJECT :
I chose to join this project because as part of the European Commission funded Marie Curie Innovative training network (ITN) it reveals to be an ambitious project that aims to obtain important results in the area of microbiology and foodborne pathogens. This project allows the integration of several vanguard techniques and in my point of view, will be followed by very good researchers. Allied to these reasons, the fact that it can broaden my international experience and the possibility to have contact with different realities of European countries makes this project ideal for future follow-up of my academic and scientific career. I was recruited for the sub-project 5 and I will be based at the Microbiology Unit at INRA-Clermont Ferrand (INRA, UR454 MICROBIOLOGIE, Centre Auvergne Rhône-Alpes, 63122 Saint Genès Champanelle ) with the supervision of Dr. Michel Hébraud.
ABOUT MY RESEARCH PROJECT :
My sub-project will connect the role of secreted protein, particularly the cell surface-associated proteins named surfaceome and the extracellular proteins named exoproteome during biofilm formation and adaptation to controlled environments (eg low air relative humidity, temperature,…). A combination of classical and innovative proteomic approaches by using different protein extraction methods (precipitation, cell fractionation, cell enzymatic shaving), in-gel and off-gel protein separation and analysis (2-DE and MALDI-TOF MS; label-free quantitative LCMS/MS) and MALDI imaging MS of proteins, peptides and lipids in biofilms will be implemented to explore the molecular response of L. monocytogenes. The analyses will be performed both with a reference sequenced strain and a strain qualified as persistent due to its identification during several years in a foodstuff plant (but never found on output food products). Deletion mutants will be constructed to understand the regulation and role of some proteins of interest. We predict difficulties when comparing our proteomic results with the transcriptomic data but we are also confident to overcome these obstacles with sub-project cooperation. We expect to reach the quantitative proteome allowing linkage between mRNA levels and protein levels in order to strengthen interpretation of transcriptome data and to decipher the molecular response in the interface cell/environment to contribute to the overall project objective of studying regulatory circuitry that drives adaptation and virulence of L. monocytogenes from farm to fork.
OTHERS INTERESTS:
Since I will be in France for the all of my PhD I intend to learn to speak and write correctly in French and to have the opportunity to visit different countries and cultures, hopefully at the same time as scientific meetings of my field.
CONTACT :
https://www.researchgate.net/profile/Tiago_Santos12
This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska Curie grant agreement n° 641984
