CEP290 tethers flagellar transition zone microtubules to the membrane and regulates flagellar protein content.
Mutations in human CEP290 cause cilia-related disorders that range in severity from isolated blindness to perinatal lethality. Here, we describe a Chlamydomonas reinhardtii mutant in which most of the CEP290 gene is deleted. Immunoelectron microscopy indicated that CEP290 is located in the flagellar transition zone in close association with the prominent microtubule-membrane links there. Ultrastructural analysis revealed defects in these microtubule-membrane connectors, resulting in loss of attachment of the flagellar membrane to the transition zone microtubules. Biochemical analysis of isolated flagella revealed that the mutant flagella have abnormal protein content, including abnormal levels of intraflagellar transport proteins and proteins associated with ciliopathies. Experiments with dikaryons showed that CEP290 at the transition zone is dynamic and undergoes rapid turnover. The results indicate that CEP290 is required to form microtubule-membrane linkers that tether the flagellar membrane to the transition zone microtubules, and is essential for controlling flagellar protein composition.
Craige
Branch
B
Department of Cell Biology, University of Massachusetts Medical School, Worcester, MA 01655, USA.
Tsao
Che-Chia
CC
Diener
Dennis R
DR
Hou
Yuqing
Y
Lechtreck
Karl-Ferdinand
KF
Rosenbaum
Joel L
JL
Witman
George B
GB
D000818:Animals; D001692:Biological Transport; D022082:Cellular Structures / Q000378:metabolism*; D016825:Chlamydomonas reinhardtii / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D002923:Cilia / Q000378:metabolism; D005407:Flagella / Q000378:metabolism / Q000502:physiology / Q000648:ultrastructure; D017353:Gene Deletion; D008566:Membranes / Q000378:metabolism; D016253:Microscopy, Immunoelectron; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism* / Q000648:ultrastructure*; D009154:Mutation
D008869:Microtubule-Associated Proteins
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