title: Crystal structure of YeaZ from Pseudomonas aeruginosa.
abstract: The Pseudomonas aeruginosa PA3685 locus encodes a conserved protein that shares 49% sequence identity with Escherichia coli YeaZ, which was recently reported as involved in the biosynthesis of threonylcarbamoyl adenosine (t(6)A), a universal modified tRNA nucleoside. Many YeaZ orthologues were reported as "essential for life" among various bacterial species, suggesting a critical role for both these proteins and for the t(6)A biosynthetic pathway. We provide here evidences that PA3685 protein (PaYeaZ) is essential. Additionally, we describe its purification, crystallization, and crystallographic structure. The crystal structure shows that PaYeaZ is composed of two domains one of which is the platform to form protein-protein interaction involved either in homodimeric assembly or in the formation of the multiprotein complex required for the synthesis of t(6)A. These features make the PaYeaZ protein a potential target candidate for the design of novel inhibitors able to hinder the complex formation and expected to abolish the crucial activity of t(6)A synthesis.
authors.affiliation: Dipartimento di Bioscienze, Università di Milano, Via Celoria 26, I-20133, Milano, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Vecchietti
authors.affiliation: Dipartimento di Bioscienze, Università di Milano, Via Celoria 26, I-20133, Milano, Italy.
authors.forename: Silvia
authors.initials: S
authors.lastname: Ferrara
authors.affiliation: Dipartimento di Bioscienze, Università di Milano, Via Celoria 26, I-20133, Milano, Italy.
authors.forename: Ruggero
authors.initials: R
authors.lastname: Rusmini
authors.affiliation: Dipartimento di Bioscienze, Università di Milano, Via Celoria 26, I-20133, Milano, Italy.
authors.forename: Raffaella
authors.initials: R
authors.lastname: Macchi
authors.affiliation: Dipartimento di Bioscienze, Università di Milano, Via Celoria 26, I-20133, Milano, Italy; CNR-Istituto di Biofisica, Via Celoria 26, I-20133, Milano, Italy. Electronic address: mario.milani@unimi.it.
authors.forename: Mario
authors.initials: M
authors.lastname: Milani
authors.affiliation: Dipartimento di Bioscienze, Università di Milano, Via Celoria 26, I-20133, Milano, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Bertoni
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002470:Cell Survival / Q000502:physiology; D003461:Crystallography; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure*; D008969:Molecular Sequence Data; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000502:physiology*; D013329:Structure-Activity Relationship
chemical_list: D029968:Escherichia coli Proteins; C512523:YeaZ protein, E coli
keywords: Crystal structure; Essential gene; Protein characterization; Pseudomonas aeruginosa; Threonylcarbamoyl adenosine synthesis