Essential role of the chaperonin CCT in rod outer segment biogenesis

Satyabrata Sinha; Marycharmain Belcastro; Poppy Datta; Seongjin Seo; Maxim Sokolov

doi:10.1167/iovs.14-13889

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Essential role of the chaperonin CCT in rod outer segment biogenesis

Journal article

Open access

Peer reviewed

Essential role of the chaperonin CCT in rod outer segment biogenesis

Satyabrata Sinha, Marycharmain Belcastro, Poppy Datta, Seongjin Seo and Maxim Sokolov

Investigative ophthalmology & visual science, Vol.55(6), pp.3775-3784

05/22/2014

DOI: 10.1167/iovs.14-13889

PMCID: PMC4062400

PMID: 24854858

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Abstract

While some evidence suggests an essential role for the chaperonin containing t-complex protein 1 (CCT) in ciliogenesis, this function remains poorly understood mechanistically. We used transgenic mice, previously generated in our lab, and characterized by a genetically-induced suppression of CCT in rod photoreceptors as well as a malformation of the rod sensory cilia, the outer segments, to gain new insights into this underlying molecular mechanism. The CCT activity in rod photoreceptors of mice was suppressed by overexpressing the chaperonin inhibitor, phosducin-like protein short, and the ensuing changes of cellular morphology were analyzed by light and electron microscopy. Protein expression levels were studied by fluorescent microscopy and Western blotting. Suppressing the chaperonin made the photoreceptors incompetent to build their outer segments. Specifically, the CCT-deficient rods appeared unable to expand the outer segment plasma membrane, and accommodate growth of this compartment. Seeking the molecular mechanisms underlying such a shortcoming, we found that the affected rods could not express normal levels of Bardet-Biedl Syndrome (BBS) proteins 2, 5, and 7 and, owing to that deficiency, were unable to assemble the BBSome, a multisubunit complex responsible for ciliary trafficking. A similar effect in response to the chaperonin suppression was also observed in cultured ciliated cells. Our data provide new evidence indicating the essential role of the chaperonin CCT in the biogenesis of vertebrate photoreceptor sensory cilia, and suggest that it may be due to the direct participation of the chaperonin in the posttranslational processing of selected BBS proteins and assembly of the BBSome.

Animals

Bardet-Biedl Syndrome - genetics

Bardet-Biedl Syndrome - metabolism

Bardet-Biedl Syndrome - pathology

Blotting, Western

Cell Line

Chaperonin Containing TCP-1 - biosynthesis

Chaperonin Containing TCP-1 - genetics

Disease Models, Animal

DNA - genetics

Electroretinography

Gene Expression Regulation, Developmental

Mice

Mice, Transgenic

Rod Cell Outer Segment - metabolism

Rod Cell Outer Segment - pathology

Details

Title: Subtitle: Essential role of the chaperonin CCT in rod outer segment biogenesis
Creators: Satyabrata Sinha - West Virginia University
Marycharmain Belcastro - West Virginia University
Poppy Datta - University of Iowa
Seongjin Seo - University of Iowa
Maxim Sokolov - West Virginia University
Resource Type: Journal article
Publication Details: Investigative ophthalmology & visual science, Vol.55(6), pp.3775-3784
DOI: 10.1167/iovs.14-13889
PMID: 24854858
PMCID: PMC4062400
ISSN: 0146-0404
eISSN: 1552-5783
Grant note: GM103488 / NIGMS NIH HHS P20 RR016477 / NCRR NIH HHS EY019665 / NEI NIH HHS R01 EY019665 / NEI NIH HHS RR016440 / NCRR NIH HHS R01 EY022616 / NEI NIH HHS P30 RR031155 / NCRR NIH HHS P20 RR016440 / NCRR NIH HHS RR031155 / NCRR NIH HHS EY022616 / NEI NIH HHS P30 RR032138 / NCRR NIH HHS P30 GM103488 / NIGMS NIH HHS
Language: English
Date published: 05/22/2014
Academic Unit: Ophthalmology and Visual Sciences
Record Identifier: 9984186903102771

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