PEAK3/C19orf35 pseudokinase, a new NFK3 kinase family member, inhibits CrkII through dimerization

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 116; no. 31; pp. 15495 - 15504
• Main Authors: Lopez, Mitchell L., Lo, Megan, Kung, Jennifer E., Dudkiewicz, Małgorzata, Jang, Gwendolyn M., Von Dollen, John, Johnson, Jeffrey R., Krogan, Nevan J., Pawłowski, Krzysztof, Jura, Natalia
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 30-07-2019
• Series: PNAS Plus
• Subjects: Actin; Amino Acid Motifs; Amino Acid Sequence; Animals; Basic Medicine; Binding; Biological Sciences; Birds; Cell and Molecular Biology; Cell Membrane - metabolism; Cell Shape; Cell size; Cell- och molekylärbiologi; Chlorocebus aethiops; Conserved Sequence; COS Cells; CrkII; Cytoskeletal Proteins - chemistry; Cytoskeletal Proteins - metabolism; Dimerization; Disruption; Elongation; Evolution, Molecular; Filaments; HEK293 Cells; Humans; Kinases; Medical and Health Sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Membrane ruffling; Motility; NKF3 family; Phylogeny; PNAS Plus; Protein Binding; Protein Domains; Protein Interaction Mapping; protein kinase; Protein Multimerization; Protein structure; Protein-Tyrosine Kinases - chemistry; Protein-Tyrosine Kinases - metabolism; Proteins; Proto-Oncogene Proteins c-crk - metabolism; pseudokinase; Regulators; Shape effects; CrkII; motility; NKF3 family; protein kinase; pseudokinase
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1906360116

Members of the New Kinase Family 3 (NKF3), PEAK1/SgK269 and Pragmin/SgK223 pseudokinases, have emerged as important regulators of cell motility and cancer progression. Here, we demonstrate that C19orf35 (PEAK3), a newly identified member of the NKF3 family, is a kinase-like protein evolutionarily conserved across mammals and birds and a regulator of cell motility. In contrast to its family members, which promote cell elongation when overexpressed in cells, PEAK3 overexpression does not have an elongating effect on cell shape but instead is associated with loss of actin filaments. Through an unbiased search for PEAK3 binding partners, we identified several regulators of cell motility, including the adaptor protein CrkII. We show that by binding to CrkII, PEAK3 prevents the formation of CrkII-dependent membrane ruffling. This function of PEAK3 is reliant upon its dimerization, which is mediated through a split helical dimerization domain conserved among all NKF3 family members. Disruption of the conserved DFG motif in the PEAK3 pseudokinase domain also interferes with its ability to dimerize and subsequently bind CrkII, suggesting that the conformation of the pseudokinase domain might play an important role in PEAK3 signaling. Hence, our data identify PEAK3 as an NKF3 family member with a unique role in cell motility driven by dimerization of its pseudokinase domain.