コンテンツへスキップ
Merck
  • Accumulation of non-outer segment proteins in the outer segment underlies photoreceptor degeneration in Bardet-Biedl syndrome.

Accumulation of non-outer segment proteins in the outer segment underlies photoreceptor degeneration in Bardet-Biedl syndrome.

Proceedings of the National Academy of Sciences of the United States of America (2015-07-29)
Poppy Datta, Chantal Allamargot, Joseph S Hudson, Emily K Andersen, Sajag Bhattarai, Arlene V Drack, Val C Sheffield, Seongjin Seo
要旨

Compartmentalization and polarized protein trafficking are essential for many cellular functions. The photoreceptor outer segment (OS) is a sensory compartment specialized for phototransduction, and it shares many features with primary cilia. As expected, mutations disrupting protein trafficking to cilia often disrupt protein trafficking to the OS and cause photoreceptor degeneration. Bardet-Biedl syndrome (BBS) is one of the ciliopathies associated with defective ciliary trafficking and photoreceptor degeneration. However, precise roles of BBS proteins in photoreceptor cells and the underlying mechanisms of photoreceptor degeneration in BBS are not well understood. Here, we show that accumulation of non-OS proteins in the OS underlies photoreceptor degeneration in BBS. Using a newly developed BBS mouse model [Leucine zipper transcription factor-like 1 (Lztfl1)/Bbs17 mutant], isolated OSs, and quantitative proteomics, we determined 138 proteins that are enriched more than threefold in BBS mutant OS. In contrast, only eight proteins showed a more than threefold reduction. We found striking accumulation of Stx3 and Stxbp1/Munc18-1 and loss of polarized localization of Prom1 within the Lztfl1 and Bbs1 mutant OS. Ultrastructural analysis revealed that large vesicles are formed in the BBS OS, disrupting the lamellar structure of the OS. Our findings suggest that accumulation (and consequent sequestration) of non-OS proteins in the OS is likely the primary cause of photoreceptor degeneration in BBS. Our data also suggest that a major function of BBS proteins in photoreceptors is to transport proteins from the OS to the cell body or to prevent entry of non-OS proteins into the OS.

材料
製品番号
ブランド
製品内容

Sigma-Aldrich
グリセロール, for molecular biology, ≥99.0%
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
塩化ナトリウム, for molecular biology, DNase, RNase, and protease, none detected, ≥99% (titration)
Sigma-Aldrich
尿素, powder, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
塩化ナトリウム 溶液, 0.9% in water, BioXtra, suitable for cell culture
Sigma-Aldrich
塩化ナトリウム, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥99%
Sigma-Aldrich
塩化ナトリウム 溶液, 5 M in H2O, BioReagent, for molecular biology, suitable for cell culture
Sigma-Aldrich
エタノール, JIS special grade, ≥99.5%
Sigma-Aldrich
エチレングリコール-ビス(2-アミノエチルエーテル)-N,N,N′,N′-四酢酸, for molecular biology, ≥97.0%
Sigma-Aldrich
尿素 溶液, BioUltra, ~8 M in H2O
Supelco
Urea, 8 M (after reconstitution with 16 mL high purity water)
Sigma-Aldrich
グリセロール 溶液, 83.5-89.5% (T)
Sigma-Aldrich
抗β-アクチン抗体, マウスモノクローナル, clone AC-15, purified from hybridoma cell culture
Sigma-Aldrich
グリセロール, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for electrophoresis, ≥99% (GC)
Sigma-Aldrich
グリセロール, BioUltra, for molecular biology, anhydrous, ≥99.5% (GC)
Sigma-Aldrich
塩化ナトリウム, JIS special grade, ≥99.5%
Sigma-Aldrich
(±)-Propylene oxide, ReagentPlus®, ≥99%
Sigma-Aldrich
塩化ナトリウム 溶液, BioUltra, for molecular biology, ~5 M in H2O
Sigma-Aldrich
塩化ナトリウム, 99.999% trace metals basis
Sigma-Aldrich
エタノール, SAJ first grade, ≥99.5%
Sigma-Aldrich
グリセロール, FCC, FG
Sigma-Aldrich
塩化ナトリウム, BioUltra, for molecular biology, ≥99.5% (AT)
Sigma-Aldrich
尿素, BioXtra, pH 7.5-9.5 (20 °C, 5 M in H2O)
Sigma-Aldrich
四酸化オスミウム 溶液, suitable for electron microscopy, 4% in H2O
Sigma-Aldrich
尿素, BioUltra, for molecular biology, 99% (T)
Sigma-Aldrich
(±)-Propylene oxide, puriss. p.a., ≥99.5% (GC)
Sigma-Aldrich
エチルアルコール(純粋), 190 proof, ACS spectrophotometric grade, 95.0%
Sigma-Aldrich
尿素, suitable for electrophoresis
Sigma-Aldrich
尿素, ACS reagent, 99.0-100.5%
Sigma-Aldrich
塩化ナトリウム, SAJ first grade, ≥99.0%