US 2017/0283852 A1 · NOVEL UBIQUITINATION SYSTEM AND THE USES THEREOF

Abstract

An unprecedented mechanism of ubiquitination that is independent of E1 and E2 enzymes, instead relying on activation of ubiquitin by ADP-ribosylation, and which is mediated by members of the SidE effector family encoded by the bacterial pathogen Legionella pneumophila is disclosed. The herein disclosed method demonstrates a method in which ubiquitination can be carried out by a single enzyme. In addition, the present disclosure also provides compositions that may be used in ubiquitination assays and/or methods of screening active substance that may inhibit the ubiquitination process.

Claims12

Dependent claims

A composition for ATP independent ubiquitination, comprising:

a) β-nicotinamide adenine dinucleotide (β-NAD);
b) a ubiquitin or a variant of the ubiquitin;
and
c) a protein selected from the group consisting of SEQ ID Nos:1-4 (SdeA, SdeB, SdeC, and SidE respectively), or a variant of any of SEQ ID Nos:
1-4, and any combination thereof, wherein the variant of SEQ ID Nos:
1-4 has at least 95% sequence identity to the corresponding SEQ ID Nos:
1-4 and maintains a putative mono-ADP-ribosyltransferase motif (R-S-ExE);
wherein the protein transfers an ADP from β-NAD to the ubiquitin or the variant of the ubiquitin to form an ADP-ribosylated ubiquitin.

5 more elements

Dependencies are inferred from the claim text.

Facts

Application

15/478,977Filed 4 Apr 2017

Priority

5 Apr 2016

Published

5 Oct 2017

Term ends

Not estimatedThe source records don’t support an estimate.

Classification10 codes

C12Q 1/48
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
C12Q 1/34
Measuring or testing processes involving enzymes, nucleic acids or microorganisms ; Compositions therefor; Processes of preparing such…
C12Q 1/37
Involving hydrolase: involving peptidase or proteinase
C12Y 204/02036
Pentosyltransferases: nAD--diphthamide ADP-ribosyltransferase

Novel ubiquitination system and the uses thereof

Life of this patent

Novel ubiquitination system and the uses thereof

Life of this patent

Abstract

Claims12