title: HrpG and HrpV proteins from the Type III secretion system of Erwinia amylovora form a stable heterodimer.
abstract: Bacterial type III secretion systems (T3SSs) are specialized multicomponent nanomachines that mediate the transport of proteins either to extracellular locations or directly into eukaryotic host cell cytoplasm. Erwinia amylovora, the main agent of rosaceous plants fireblight disease, employs an Hrp/Hrc1 T3SS to accomplish its pathogenesis. The regulatory network that controls the activation of this T3SS is largely unknown in E. amylovora. However, in Pseudomonas syringae pathovars, the HrpG/HrpV complex has been shown to directly regulate the activity of transcription factor HrpS and consequently the upregulation of the Hrp/Hrc1 T3SS related genes. In this work, we report the successful recombinant production and purification of a stable E. amylovora HrpG/HrpV complex, using pPROpET, a bicistronic expression vector. Furthermore, we present the first solution structure of this complex based on small-angle X-ray scattering data.
authors: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas (IMBB-FORTH), Nikolaou Plastira 100, GR-70013, Heraklion, Crete, Greece Biology Department, University of Crete, Vasilika Vouton, 71409 Heraklio, Crete, Greece gazi@lebs.cnrs-gif.fr anadegaz@gmail.com.
authors: Anastasia D
authors: AD
authors: Gazi
authors: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas (IMBB-FORTH), Nikolaou Plastira 100, GR-70013, Heraklion, Crete, Greece.
authors: Spyridoula
authors: S
authors: Charova
authors: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas (IMBB-FORTH), Nikolaou Plastira 100, GR-70013, Heraklion, Crete, Greece.
authors: Michalis
authors: M
authors: Aivaliotis
authors: Professor Emeritus, Biology Department, University of Crete, Vasilika Vouton, 71409 Heraklio, Crete, Greece Professor Emeritus, Department of Environmental Sciences, Policy, and Management, University of California, Berkeley, 130 Mulford Hall, Berkeley, CA 94720-3114, USA.
authors: Nicholas J
authors: NJ
authors: Panopoulos
authors: Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas (IMBB-FORTH), Nikolaou Plastira 100, GR-70013, Heraklion, Crete, Greece Biology Department, University of Crete, Vasilika Vouton, 71409 Heraklio, Crete, Greece.
authors: Michael
authors: M
authors: Kokkinidis
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D058947:Bacterial Secretion Systems; D003001:Cloning, Molecular; D044084:Erwinia amylovora / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D015870:Gene Expression; D005822:Genetic Vectors; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D053838:Scattering, Small Angle; D017386:Sequence Homology, Amino Acid
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D011994:Recombinant Proteins
keywords: Erwinia amylovora; Pseudomonas syringae; bicistronic expression vectors; hypersensitive response and pathogenicity; small-angle X-ray scattering (SAXS)