title: Proteomic analysis of isolated ciliary transition zones reveals the presence of ESCRT proteins.
abstract: The transition zone (TZ) is a specialized region of the cilium characterized by Y-shaped connectors between the microtubules of the ciliary axoneme and the ciliary membrane [1]. Located near the base of the cilium, the TZ is in the prime location to act as a gate for proteins into and out of the ciliary compartment, a role supported by experimental evidence [2-6]. The importance of the TZ has been underscored by studies showing that mutations affecting proteins located in the TZ result in cilia-related diseases, or ciliopathies, presenting symptoms including renal cysts, retinal degeneration, and situs inversus [7-9]. Some TZ proteins have been identified and shown to interact with each other through coprecipitation studies in vertebrate cells [4, 10, 11] and genetics studies in C. elegans [3]. As a distinct approach to identify TZ proteins, we have taken advantage of the biology of Chlamydomonas to isolate TZs. Proteomic analysis identified 115 proteins, ten of which were known TZ proteins related to ciliopathies, indicating that the preparation was highly enriched for TZs. Interestingly, six proteins of the endosomal sorting complexes required for transport (ESCRT) were also associated with the TZs. Identification of these and other proteins in the TZ will provide new insights into functions of the TZ, as well as candidate ciliopathy genes.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06511, USA. Electronic address: dennis.diener@yale.edu.
authors.forename: Dennis R
authors.initials: DR
authors.lastname: Diener
authors.affiliation: Department of Life Sciences, University of Siena, Siena 53100, Italy.
authors.forename: Pietro
authors.initials: P
authors.lastname: Lupetti
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06511, USA. Electronic address: joel.rosenbaum@yale.edu.
authors.forename: Joel L
authors.initials: JL
authors.lastname: Rosenbaum
mesh_terms: D002696:Chlamydomonas / Q000235:genetics / Q000378:metabolism*; D002923:Cilia / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism*; D008854:Microscopy, Electron; D028861:Nuclear Pore Complex Proteins / Q000378:metabolism; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics / Q000379:methods
chemical_list: D056827:Endosomal Sorting Complexes Required for Transport; D028861:Nuclear Pore Complex Proteins; D020543:Proteome