title: Characterization of bud emergence 46 (BEM46) protein: sequence, structural, phylogenetic and subcellular localization analyses.
abstract: The bud emergence 46 (BEM46) protein from Neurospora crassa belongs to the α/β-hydrolase superfamily. Recently, we have reported that the BEM46 protein is localized in the perinuclear ER and also forms spots close by the plasma membrane. The protein appears to be required for cell type-specific polarity formation in N. crassa. Furthermore, initial studies suggested that the BEM46 amino acid sequence is conserved in eukaryotes and is considered to be one of the widespread conserved "known unknown" eukaryotic genes. This warrants for a comprehensive phylogenetic analysis of this superfamily to unravel origin and molecular evolution of these genes in different eukaryotes. Herein, we observe that all eukaryotes have at least a single copy of a bem46 ortholog. Upon scanning of these proteins in various genomes, we find that there are expansions leading into several paralogs in vertebrates. Usingcomparative genomic analyses, we identified insertion/deletions (indels) in the conserved domain of BEM46 protein, which allow to differentiate fungal classes such as ascomycetes from basidiomycetes. We also find that exonic indels are able to differentiate BEM46 homologs of different eukaryotic lineage. Furthermore, we unravel that BEM46 protein from N. crassa possess a novel endoplasmic-retention signal (PEKK) using GFP-fusion tagging experiments. We propose that three residues namely a serine 188S, a histidine 292H and an aspartic acid 262D are most critical residues, forming a catalytic triad in BEM46 protein from N. crassa. We carried out a comprehensive study on bem46 genes from a molecular evolution perspective with combination of functional analyses. The evolutionary history of BEM46 proteins is characterized by exonic indels in lineage specific manner.
authors.affiliation: Abteilung für Botanik mit Schwerpunkt Genetik und Molekularbiologie, Botanisches Institut und Botanischer Garten, Christian-Albrechts-Universität zu Kiel, Kiel, Germany.
authors.forename: Abhishek
authors.initials: A
authors.lastname: Kumar
authors.forename: Krisztina
authors.initials: K
authors.lastname: Kollath-Leiß
authors.forename: Frank
authors.initials: F
authors.lastname: Kempken
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004721:Endoplasmic Reticulum / Q000201:enzymology; D019143:Evolution, Molecular; D005656:Fungal Proteins / Q000737:chemistry* / Q000235:genetics; D006867:Hydrolases / Q000737:chemistry* / Q000235:genetics; D054643:INDEL Mutation; D009492:Neurospora crassa / Q000235:genetics*; D010802:Phylogeny; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary
chemical_list: D005656:Fungal Proteins; D006867:Hydrolases
keywords: Bud emergence 46 (bem46); Endoplasmic retention signal; Evolution; Fungi; Indels; eGFP-fusion; α/β-hydrolase