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Analysis of microtubule plus-end-tracking proteins in cilia.
- Source :
-
Methods in enzymology [Methods Enzymol] 2013; Vol. 524, pp. 105-22. - Publication Year :
- 2013
-
Abstract
- The microtubule (MT) plus-end-tracking proteins (+TIPs) belonging to the end binding (EB) protein family have been studied extensively in the context of cytoplasmic MTs and were shown to play pivotal roles in regulating MT dynamics and in recruiting other +TIPs to MT ends. Early studies in the green alga Chlamydomonas reinhardtii showed that EB1 localizes to the distal flagellum tip and the basal body, and subsequent studies using green fluorescent protein-tagged fusion proteins have demonstrated similar localization of EBs in other ciliated organisms, including mammalian cells as demonstrated here. Functional analysis of EBs in cultured mammalian cells revealed that EB1 and EB3 are required for biogenesis of primary cilia. Although mammalian EB3 localizes to the tip of some cilia and induces cilium elongation, EBs primarily seem to promote ciliogenesis via MT minus-end anchoring at the basal body, in turn facilitating vesicular trafficking to the cilium base. Moreover, mammalian EBs were shown to interact with several proteins implicated in MT minus-end anchoring and/or vesicular trafficking to cilia. Recent work suggests that apart from EBs, additional +TIPs may be present at the distal tip of cilia where they could regulate axoneme assembly, stability, or disassembly.<br /> (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Subjects :
- Cells, Cultured
Chlamydomonas reinhardtii chemistry
Cilia chemistry
Epithelial Cells chemistry
Fibroblasts chemistry
Flagella chemistry
Gene Expression
Green Fluorescent Proteins antagonists & inhibitors
Green Fluorescent Proteins genetics
Green Fluorescent Proteins metabolism
Humans
Microscopy, Fluorescence
Microtubule-Associated Proteins antagonists & inhibitors
Microtubule-Associated Proteins genetics
Microtubules
Molecular Imaging
RNA, Small Interfering genetics
Recombinant Fusion Proteins antagonists & inhibitors
Recombinant Fusion Proteins genetics
Recombinant Fusion Proteins metabolism
Chlamydomonas reinhardtii metabolism
Cilia metabolism
Epithelial Cells metabolism
Fibroblasts metabolism
Flagella metabolism
Microtubule-Associated Proteins metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1557-7988
- Volume :
- 524
- Database :
- MEDLINE
- Journal :
- Methods in enzymology
- Publication Type :
- Academic Journal
- Accession number :
- 23498737
- Full Text :
- https://doi.org/10.1016/B978-0-12-397945-2.00007-X