Divergent kinase WNG1 is regulated by phosphorylation of an atypical activation sub-domain

Pravin S. Dewangan, Tsebaot G. Beraki, E. Ariana Paiz, Delia Appiah Mensah, Zhe Chen, Michael L. Reese

Research output: Contribution to journalArticlepeer-review

Abstract

Apicomplexan parasites like Toxoplasma gondii grow and replicate within a specialized organelle called the parasitophorous vacuole. The vacuole is decorated with parasite proteins that integrate into the membrane after trafficking through the parasite secretory system as soluble, chaperoned complexes. A regulator of this process is an atypical protein kinase called WNG1. Phosphorylation by WNG1 appears to serve as a switch for membrane integration. However, like its substrates, WNG1 is secreted from the parasite dense granules, and its activity must, therefore, be tightly regulated until the correct membrane is encountered. Here, we demonstrate that, while another member of the WNG family can adopt multiple multimeric states, WNG1 is monomeric and therefore not regulated by multimerization. Instead, we identify two phosphosites on WNG1 that are required for its kinase activity. Using a combination of in vitro biochemistry and structural modeling, we identify basic residues that are also required for WNG1 activity and appear to recognize the activating phosphosites. Among these coordinating residues are the 'HRD' Arg, which recognizes activation loop phosphorylation in canonical kinases. WNG1, however, is not phosphorylated on its activation loop, but rather on atypical phosphosites on its C-lobe. We propose a simple model in which WNG1 is activated by increasing ATP concentration above a critical threshold once the kinase traffics to the parasitophorous vacuole.

Original languageEnglish (US)
Pages (from-to)1877-1889
Number of pages13
JournalThe Biochemical journal
Volume479
Issue number17
DOIs
StatePublished - Sep 16 2022

Keywords

  • apicomplexa
  • cellular secretion
  • phosphorylation
  • pseudokinases

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Divergent kinase WNG1 is regulated by phosphorylation of an atypical activation sub-domain'. Together they form a unique fingerprint.

Cite this