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PhD Position in Mechanisms of Amyloid Aggregate Sorting in Yeast
Category:
Phd

Description:

Dr. J. Tyedmers conducts research in a bridging position between the “Krehl Klinik” and the “ZMBH”. The laboratory is located in the Department of Prof. Dr. P. Nawroth, but he is also affiliated with the laboratory of Prof. Dr. Bernd Bukau that is interested in various aspects of protein folding and protein quality control.

Research Area: The formation of protein aggregates including amyloids is a hallmark of several neurodegenerative diseases, diabetes and cellular aging, amongst others. A commonly employed model organism to study protein aggregation, Saccharomyces cerevisiae, possesses at least three independent Protein Quality Control Sites (PQC’s) where different types of aggregates are deposited. The group of J. Tyedmers studies one of these deposition sites termed “Insoluble Protein Deposit (IPOD)” and has most recently discovered that amyloid aggregates are recruited to the IPOD via vesicular transport along actin tracks. The project aims 1) to characterize the molecular composition of this novel recruitment machinery, 2) to determine the physiological role of the IPOD and its influence on cellular homeostasis, 3) to study the fate of aggregates deposited at the IPOD.

In the Ph.D. project, you will use different IPOD substrates fused to GFP to 1) screen for novel factors that contribute to the recruitment of amyloid aggregates to the IPOD; 2) purify IPODs and transport intermediates on transit to this site using a Fluorescence Activated Cell Sorting (FACS)-based method to identify additional substrates and structural components of the IPOD; 3) further characterize the novel recruitment machinery using fluorescence microscopy and biochemical and genetic yeast methods.

References:

• Kumar R, Nawroth P, Tyedmers J. Prion Aggregates are recruited to the Insoluble Protein Deposit (IPOD) via Myosin 2-based vesicular transport. PLoS Genet. 2016 Sep 30;12(9):e1006324

• Hernandez-Hidalgo I, Fleming T, Eckstein V, Herzig S, Nawroth P, Tyedmers J. Characterization of Aggregate Load & Pattern in living Yeast Cells by Flow Cytometry. Biotechniques. 2016 Sep 1;61(3):137-48.

• Tyedmers J. Patterns of [PSI+] aggregation allow insights into cellular organization of yeast prion aggregates. Prion. 2012 Jul 1;6(3):191-200.

• Saibil HR, Seybert A, Habermann A, Winkler J, Eltsov M, Perkovic M, Castaño-Diez D, Scheffer MP, Haselmann U, Chlanda P, Lindquist S, Tyedmers J, Frangakis AS. Heritable yeast prions have a highly organized 3-dimensional architecture with inter-fiber structures. Proc Natl Acad Sci U S A. 2012 Sep 11;109(37):14906-11.

• Tyedmers J, Mogk, A & Bukau, B. Cellular strategies for controlling protein aggregation. Nat Rev Mol Cell Biol. 2010 Nov;11(11):777-88

• Tyedmers J, Treusch S, Dong J, McCaffery JM, Bevis B, Lindquist S. Prion induction involves an ancient system for the sequestration of aggregated proteins and heritable changes in prion fragmentation. Proc Natl Acad Sci U S A. 2010 May 11;107(19):8633-8.

Methods that will be used:

• general yeast genetics methods

• standard molecular biology techniques

• fluorescence microscopy

• Screening with yeast libraries

• Flow Cytometry

• FACS-based isolation strategies for aggregates

Cooperation partners:

Prof H. Saibil, London

Prof. B. Bukau, Heidelberg

Prof. P. Nawroth, Heidelberg

Personal qualifications:

We are looking for a highly motivated and communicative Ph.D. student who is interested to perform competitive high quality scientific research in the area of protein misfolding and aggregation. She or he will have a Masters Degree in Life sciences acceptable to HBIGS and a strong background in molecular biology and/or cell biology. Previous experience in working with yeast, High-throughput screening and/or Flow Cytometry would be an advantage.

Keywords:

Neurodegenerative Disease, Prions, Saccharomyces cerevisiae, Protein Aggregation, Amyloid Aggregates, Protein Quality Control, Insoluble Protein Deposit (IPOD), Vesicular Transport