US 5,705,387 A · Heat-inducible N-degron module

Abstract

The N-degron is an intracellular degradation signal whose essential determinant is a specific, destabilizing, N-terminal amino acid residue. A set of N-degrons containing different destabilizing residues is manifested as the N-end rule, which relates the in vivo half-life of a protein to the identity of its N-terminal amino acid residue. Disclosed herein is a heat-inducible N-degron module. A heat-inducible N-degron module is a protein or peptide bearing a destabilizing N-terminal amino acid residue which becomes a substrate of the N-end rule pathway only at a temperature high enough to result in at least partial unfolding of the protein. At this elevated (nonpermissive) temperature, the heat-inducible N-degron module (and any protein or peptide attached at its C-terminus) is rapidly degraded in a cell in which the N-end rule pathway is operative. Also disclosed are DNA and protein fusion constructs, methods for screening for additional heat-inducible N-degron modules and methods for using the disclosed heat-inducible N-degron modules.

Claims9

Dependent claims

A heat-inducible N-degron module comprising:

a) a destabilizing N-terminal amino acid residue;
b) an internal lysine amino acid residue which functions as an attachment site for a multiubiquitin chain at a permissive temperature, but not at a non-permissive temperature in a eukaryotic cell or cell extract;
the heat-inducible N-degron module being encoded by a DNA sequence which hybridizes, under stringent hybridization conditions, to the DNA sequence represented in SEQ ID NO:
1.

1 more element

Dependencies are inferred from the claim text.

Heat-inducible N-degron module

Life of this patent

Heat-inducible N-degron module

Life of this patent

Abstract

Claims9