Threonine eliminylation by bacterial phosphothreonine lyases rapidly causes cross-linking of mitogen-activated protein kinase (MAPK) in live cells.

Abstract

Old long-lived proteins contain dehydroalanine (Dha) and dehydrobutyrine (Dhb), two amino acids engendered by dehydration of serines and threonines, respectively. Although these residues have a suspected role in protein cross-linking and aggregation, their direct implication has yet to be determined. Here, we have taken advantage of the ability of the enteropathogen Shigella to convert the phosphothreonine residue of the pT-X-pY consensus sequence of ERK and p38 into Dhb and followed the impact of dehydration on the fate of the two MAPKs. To that end, we have generated the first antibodies recognizing Dhb-modified proteins and allowing tracing them as they form. We showed that Dhb modifications accumulate in a long-lasting manner in Shigella-infected cells, causing subsequent formation of covalent cross-links of MAPKs. Moreover, the Dhb signal correlates precisely with the activation of the Shigella type III secretion apparatus, thus evidencing injectisome activity. This observation is the first to document a causal link between Dhb formation and protein cross-linking in live cells. Detection of eliminylation is a new avenue to phosphoproteome regulation in eukaryotes that will be instrumental for the development of type III secretion inhibitors.

• Authors: Benoit M Meijer, Suk Min Jang, Ida C Guerrera, Cerina Chhuon, Joanna Lipecka, Caroline Reisacher, Françoise Baleux, Philippe J Sansonetti, Christian Muchardt, Laurence Arbibe
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 292 Issue 19 Pg. 7784-7794 (05 12 2017) ISSN: 1083-351X [Electronic] United States
• PMID: 28325837 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

Chemical References

• Aminobutyrates
• Antibodies
• Bacterial Proteins
• Type III Secretion Systems
• dehydrobutyrine
• Threonine
• dehydroalanine
• Extracellular Signal-Regulated MAP Kinases
• p38 Mitogen-Activated Protein Kinases
• Carbon-Oxygen Lyases
• Alanine

Topics

• Alanine (analogs & derivatives, chemistry)
• Aminobutyrates (chemistry)
• Animals
• Antibodies (chemistry)
• Bacterial Proteins (metabolism)
• Caco-2 Cells
• Carbon-Oxygen Lyases (metabolism)
• Cell Line
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• HeLa Cells
• Humans
• MAP Kinase Signaling System
• Mice
• Protein Binding
• Proteomics
• Shigella (enzymology)
• Substrate Specificity
• Threonine (chemistry)
• Type III Secretion Systems
• p38 Mitogen-Activated Protein Kinases (metabolism)