Journal Article

Deciphering the evolution of composite-type GSKIP in mitochondria and Wnt signaling pathways

Lin, Chen-Yen, Kwan, Aij-Li, Cheng, Yu-Fan, Lin, Sin-Yi, Cheng, Jiin-Tsuey, Ko, Huey-Jiun, Tsai, Cheng-Yu, Loh, Joon-Khim, Chiou, Shean-Jaw, Lai, Yun-Ling, Liu, Hsin-Fu, Wang, Chihuei, Hong, Yi-Ren

2022

Overview

We previously revealed the origin of mammalian simple-type glycogen synthase kinase interaction protein (GSKIP), which served as a scavenger and a competitor in the Wnt signaling pathway during evolution. In this study, we investigated the conserved and nonconserved regions of the composite-type GSKIP by utilizing bioinformatics tools, site-directed mutagenesis, and yeast two-hybrid methods. The regions were denoted as the pre-GSK3β binding site, which is located at the front of GSK3β-binding sites. Our data demonstrated that clustered mitochondria protein 1 (CLU1), a type of composite-type GSKIP that exists in the mitochondria of all eukaryotic organisms, possesses the protein known as domain of unknown function 727 (DUF727), with a pre-GSK3β-binding site and a mutant GSK3β-binding flanking region. Another type of composite-type GSKIP, armadillo repeat containing 4 (ARMC4), which is known for cilium movement in vertebrates, contains an unintegrated DUF727 flanking region with a pre-GSK3β-binding site (115SPxF118) only. In addition, the sequence of the GSK3β-binding site in CLU1 revealed that Q126L and V130L were not conserved, differing from the ideal GSK3β-binding sequence of simple-type GSKIP. We further illustrated two exceptions, namely 70 kilodalton heat shock proteins (Hsp70/DnaK) and Mitofilin in nematodes, that presented an unexpected ideal GSK3β-binding region with a pre-GSK3β sequence; this composite-type GSKIP could only occur in vertebrate species. Furthermore, we revealed the importance of the pre-GSK3β-binding site (118F or 118Y) and various mutant GSK3β-binding sites of composite-type GSKIP. Collectively, our data suggest that the new composite-type GSKIP starts with a DUF727 domain followed by a pre-GSK3β-binding site, with the subsequent addition of the GSK3β-binding site, which plays vital roles for CLU1, Mitofilin, and ARMC4 in mitochondria and Wnt signaling pathways during evolution.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Alzheimer's disease / Alzheimers disease / Amino Acid Sequence / Amino acids / Analysis / Animals / Armadillo Domain Proteins - chemistry / Armadillo Domain Proteins - genetics / Armadillo Domain Proteins - metabolism / Binding Sites / Binding sites (Biochemistry) / Biochemistry / Bioinformatics / Biology and Life Sciences / Cloning, Molecular / Computer and Information Sciences / Conserved Sequence / Data mining / DnaK protein / Domains / Evolution / Evolution, Molecular / Glycogen / Glycogen synthase kinase 3 / Glycogen Synthase Kinase 3 beta - metabolism / Glycogens / Heat shock proteins / Hospitals / Hsp70 protein / Humans / Kinases / Lung cancer / Maximum likelihood method / Medical research / Medical schools / Medicine / Mitochondria / Mitochondria - metabolism / Models, Molecular / Mutagenesis / Mutagenesis, Site-Directed / Mutants / Nematodes / Neurosurgery / Ontology / Pathways / Peptides / Phosphorylation / Phylogenetics / Phylogeny / Protein Binding / Protein Conformation / Proteins / Repressor Proteins - chemistry / Repressor Proteins - genetics / Repressor Proteins - metabolism / Research and Analysis Methods / RNA-Binding Proteins - chemistry / RNA-Binding Proteins - genetics / RNA-Binding Proteins - metabolism / Sequence Analysis, DNA / Signal transduction / Signaling / Site-directed mutagenesis / Software / Two-Hybrid System Techniques / Vertebrates / Wnt protein / Wnt proteins / Wnt Signaling Pathway / Yeast / Yeasts