title: The Chlamydia effector chlamydial outer protein N (CopN) sequesters tubulin and prevents microtubule assembly.
abstract: Chlamydia species are obligate intracellular pathogens that utilize a type three secretion system to manipulate host cell processes. Genetic manipulations are currently not possible in Chlamydia, necessitating study of effector proteins in heterologous expression systems and severely complicating efforts to relate molecular strategies used by Chlamydia to the biochemical activities of effector proteins. CopN is a chlamydial type three secretion effector that is essential for virulence. Heterologous expression of CopN in cells results in loss of microtubule spindles and metaphase plate formation and causes mitotic arrest. CopN is a multidomain protein with similarity to type three secretion system "plug" proteins from other organisms but has functionally diverged such that it also functions as an effector protein. We show that CopN binds directly to αβ-tubulin but not to microtubules (MTs). Furthermore, CopN inhibits tubulin polymerization by sequestering free αβ-tubulin, similar to one of the mechanisms utilized by stathmin. Although CopN and stathmin share no detectable sequence identity, both influence MT formation by sequestration of αβ-tubulin. CopN displaces stathmin from preformed stathmin-tubulin complexes, indicating that the proteins bind overlapping sites on tubulin. CopN is the first bacterial effector shown to disrupt MT formation directly. This recognition affords a mechanistic understanding of a strategy Chlamydia species use to manipulate the host cell cycle.
authors.affiliation: Division of Chemical and Physical Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232, USA.
authors.forename: Tara L
authors.initials: TL
authors.lastname: Archuleta
authors.forename: Yaqing
authors.initials: Y
authors.lastname: Du
authors.forename: Chauca A
authors.initials: CA
authors.lastname: English
authors.forename: Stephen
authors.initials: S
authors.lastname: Lory
authors.forename: Cammie
authors.initials: C
authors.lastname: Lesser
authors.forename: Melanie D
authors.initials: MD
authors.lastname: Ohi
authors.forename: Ryoma
authors.initials: R
authors.lastname: Ohi
authors.forename: Benjamin W
authors.initials: BW
authors.lastname: Spiller
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D002417:Cattle; D002689:Chlamydia / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000472:pathogenicity*; D002690:Chlamydia Infections / Q000235:genetics / Q000378:metabolism; D008677:Metaphase; D008870:Microtubules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008941:Spindle Apparatus / Q000235:genetics / Q000378:metabolism; D014404:Tubulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D037521:Virulence Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D014404:Tubulin; D037521:Virulence Factors