Research highlights

past and present

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19. May 2012

How to track the ageing of proteins

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Chris Fit

+ tandem fluorescent timer proteins (tFTs)

"measure protein age and protein turnover  in space and time"

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+ Protein quality control in the nucleus 

Removal of mis-targeted proteins from the inner nuclear membrane

We identified a protein degradation pathway at the INM in yeast mediated by the Asi complex. The Asi complex functions together with the ubiquitin-conjugating enzymes Ubc6 and Ubc7 to degrade soluble and integral membrane proteins. Genetic evidence suggests that the Asi ubiquitin ligase defines a pathway distinct from, but complementary to, ERAD. Using unbiased screening with a novel genome-wide yeast library based on a tandem fluorescent protein timer, we identify more than 50 substrates of the Asi, Hrd1 and Doa10 E3 ubiquitin ligases. We show that the Asi ubiquitin ligase is involved in degradation of mislocalized integral membrane proteins, thus acting to maintain and safeguard the identity of the INM.

Khmelinskii A, Blaszczak E, Pantazopoulou M, Fischer B, Omnus DJ, Le Dez G, Brossard A, Gunnarsson A, Barry JD, Meurer M, Kirrmaier D, Boone C, Huber W, Rabut G, Ljungdahl PO & Knop M (2014) Protein quality control at the inner nuclear membrane. Nature 516: 410–413

+ N-end rule pathway  

Systematic peptide profiling reveales the logic of N-degrons

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+ Libraries for high throughput yeast cell biology

  • Arrayed libraries: SWAT 

  • Pooled libraries: CASTLING

+ Light sheet microscopes

live cell imaging without bleaching (almost)

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  • The πSPIM microscope

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  • The light pad microscope

© 2019 By Michael Knop.