Cathepsin proteases mediate photoreceptor cell degeneration in Drosophila
Department of Biology, Dartmouth College, 54 College St, Hanover, NH 03755, USA. Neurobiology of Disease
(Impact Factor: 5.08).
03/2012; 46(3):655-62. DOI: 10.1016/j.nbd.2012.03.004
Endocytosis-mediated cell death is a form of degeneration displayed in several Drosophila mutants. This form of degeneration is displayed in several Drosophila mutant lines including flies lacking the eye-specific PLC (norpA). The cell death pathway is initiated by the stabilization of complexes between rhodopsin and arrestin which undergo massive endocytosis into the cell body. The internalized rhodopsin becomes insoluble and builds up in the late endosomal system, wherein it triggers cell death. Cathepsins are resident late endosome/lysosome proteases that have been shown to mediate apoptosis in many disease models. Therefore we sought to test the involvement of cathepsins in endocytosis-mediated retinal degeneration. Here we show that cathepsins mediate cell death in light-exposed norpA eyes. Moreover, we show that the cathepsin L-like cysteine protease, CP1, specifically mediates retinal degeneration, while the aspartyl protease, cathepsin D, does not. Furthermore, eye-specific expression of pan-cathepsin inhibitors also blocks cell death. Western blot analysis demonstrates that cathepsin L levels remain unchanged during retinal degeneration. However, whole mount immunohistochemistry performed on light-exposed retinas revealed a decrease in cathepsin L levels and a loss of rhodopsin/ CP1 colocalization, suggesting that cathepsin L translocates during the degeneration process. Lastly, we show that the retinal degeneration can be enhanced by the overexpression of cathepsin L in the sensitized norpA background. Together these data show that cathepsins play a crucial role in endocytosis-mediated retinal degeneration and are consistent with a model where rhodopsin internalization and accumulation in the endosomal/lysosomal system triggers cathepsin translocation to the cytosol.
Available from: Rohit Farmer
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ABSTRACT: Cathepsin L is a cysteine protease which degrades connective tissue proteins including collagen, elastin, and fibronectin. In this study, five well-characterized cathepsin L proteins from different arthropods were used as query sequences for the Drosophila genome database. The search yielded 10 cathepsin L-like sequences, of which eight putatively represent novel cathepsin L-like proteins. To understand the phylogenetic relationship among these cathepsin L-like proteins, a phylogenetic tree was constructed based on their sequences. In addition, models of the tertiary structures of cathepsin L were constructed using homology modeling methods and subjected to molecular dynamics simulations to obtain reasonable structure to understand its dynamical behavior. Our findings demonstrate that all of the potential Drosophila cathepsin L-like proteins contain at least one cathepsin propeptide inhibitor domain. Multiple sequence alignment and homology models clearly highlight the conservation of active site residues, disulfide bonds, and amino acid residues critical for inhibitor binding. Furthermore, comparative modeling indicates that the sequence/structure/function profiles and active site architectures are conserved.
Journal of biomolecular Structure & Dynamics 12/2012; 31(12). DOI:10.1080/07391102.2012.745379 · 2.92 Impact Factor
Available from: Marja Jaattela
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ABSTRACT: Lysosomes serve as the cellular recycling centre and are filled with numerous hydrolases that can degrade most cellular macromolecules. Lysosomal membrane permeabilization and the consequent leakage of the lysosomal content into the cytosol leads to so-called "lysosomal cell death". This form of cell death is mainly carried out by the lysosomal cathepsin proteases and can have necrotic, apoptotic or apoptosis-like features depending on the extent of the leakage and the cellular context. This article summarizes our current knowledge on lysosomal cell death with an emphasis on the upstream mechanisms that lead to lysosomal membrane permeabilization.
Journal of Cell Science 05/2013; 126(Pt 9):1905-1912. DOI:10.1242/jcs.091181 · 5.43 Impact Factor
Available from: plosone.org
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ABSTRACT: Author Summary
Malfunctioning of phototransduction is the major cause of human blindness. Without functional phototransduction, rhodopsin-1, the major visual pigment, is rapidly endocytosed and accumulated in late endosomes. Impaired lysosomal delivery of endocytosed rhodopsin and its degradation has been reported to trigger progressive and light-dependent retinal degeneration in Drosophila models. It is intriguing why endocytosed rhodopsin accumulates in late endosomes instead of being delivered to lysosomes for degradation. Is this attributable to a saturation of rhodopsin endocytosis, which impedes the delivery capacity of the cell? To investigate the underlying mechanisms of rhodopsin accumulation in late endosomes, we used a suppressor of phototransduction mutants, which was identified previously from our unbiased genetic screen. This suppressor, called diehard4, shifts the membrane balance between late endosomes and lysosomes, resulting in the facilitated degradation of endocytosed rhodopsin. Our results clearly demonstrate that a previously unknown mechanism of negative regulation is actively engaged in vesicular traffic between endosomes and lysosomes in fly photoreceptors. We showed that eliminating such blockage alone was enough to rescue retinal degeneration in phototransduction mutants. From these results, we anticipate that the identification of additional components and an in-depth description of this molecular machinery will aid in therapeutic interventions of various retinal dystrophies and neurodegenerative disorders.
PLoS Genetics 06/2013; 9(6):e1003559. DOI:10.1371/journal.pgen.1003559 · 7.53 Impact Factor
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