Analysis of Intracellular Distribution and Trafficking of the CLN3 Protein in Fusion with the Green Fluorescent Proteinin Vitro

Kida, E; Kaczmarski, Wojciech; Golabek, Adam A.; Kaczmarski, A.; Michalewski, Martin P.; Wisniewski, K. E.

doi:10.1006/mgme.1999.2837

Cited by 39 publications

(28 citation statements)

References 30 publications

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“…The CLN3 protein bears consensus sites for N-glycosylation, phosphorylation and C-terminal lipid modification, including myristoylation, palmitoylation and prenylation; experimental data validates these predictions [29,42–45]. These protein modifications most likely modulate CLN3 protein stability and regulate function.…”

Section: Cln3 Mutations In Jnclmentioning

confidence: 83%

The juvenile Batten disease protein, CLN3, and its role in regulating anterograde and retrograde post-Golgi trafficking

Cotman

Staropoli

2012

Clinical Lipidology

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Loss-of-function mutations in CLN3 are responsible for juvenile-onset neuronal ceroid lipofuscinosis (JNCL), or Batten disease, which is an incurable lysosomal disease that manifests with vision loss, followed by seizures and progressive neurodegeneration, robbing children of motor skills, speech and cognition, and eventually leading to death in the second or third decade of life. Emerging clinical evidence points to JNCL pathology outside of the CNS, including the cardiovascular system. The CLN3 gene encodes an unusual transmembrane protein, CLN3 or battenin, whose elusive function has been the subject of intense study for more than 10 years. Owing to the detailed characterization of a large number of disease models, our knowledge of CLN3 protein function is finally coming into focus. This review will describe the most current understanding of CLN3 structure, function and dysfunction in JNCL.

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Section: Cln3 Mutations In Jnclmentioning

confidence: 83%

The juvenile Batten disease protein, CLN3, and its role in regulating anterograde and retrograde post-Golgi trafficking

Cotman

Staropoli

2012

Clinical Lipidology

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“…The C-terminus is farnesylated [9], [10], and this modification was found to be important for CLN3 localization in one study [8]. There are three lysosomal localization motifs in the sequence of CLN3: two dileucine sorting motifs in the cytosolic internal loop, and an acidic patch found in the C-terminus [11]–[13]. CLN3 may also be phosphorylated at serine and threonine residues [14]–[16].…”

Section: Introductionmentioning

confidence: 96%

Osmotic Stress Changes the Expression and Subcellular Localization of the Batten Disease Protein CLN3

et al. 2013

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Juvenile CLN3 disease (formerly known as juvenile neuronal ceroid lipofuscinosis) is a fatal childhood neurodegenerative disorder caused by mutations in the CLN3 gene. CLN3 encodes a putative lysosomal transmembrane protein with unknown function. Previous cell culture studies using CLN3-overexpressing vectors and/or anti-CLN3 antibodies with questionable specificity have also localized CLN3 in cellular structures other than lysosomes. Osmoregulation of the mouse Cln3 mRNA level in kidney cells was recently reported. To clarify the subcellular localization of the CLN3 protein and to investigate if human CLN3 expression and localization is affected by osmotic changes we generated a stably transfected BHK (baby hamster kidney) cell line that expresses a moderate level of myc-tagged human CLN3 under the control of the human ubiquitin C promoter. Hyperosmolarity (800 mOsm), achieved by either NaCl/urea or sucrose, dramatically increased the mRNA and protein levels of CLN3 as determined by quantitative real-time PCR and Western blotting. Under isotonic conditions (300 mOsm), human CLN3 was found in a punctate vesicular pattern surrounding the nucleus with prominent Golgi and lysosomal localizations. CLN3-positive early endosomes, late endosomes and cholesterol/sphingolipid-enriched plasma membrane microdomain caveolae were also observed. Increasing the osmolarity of the culture medium to 800 mOsm extended CLN3 distribution away from the perinuclear region and enhanced the lysosomal localization of CLN3. Our results reveal that CLN3 has multiple subcellular localizations within the cell, which, together with its expression, prominently change following osmotic stress. These data suggest that CLN3 is involved in the response and adaptation to cellular stress.

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“…The low abundance of CLN3 protein in mammalian cells has contributed to difficulties in establishing its intracellular distribution. However, the consensus view based on various studies to localize the endogenous and exogenously expressed protein is that CLN3 is most likely a lysosomal/endosomal protein trafficking through the ER and Golgi (Järvelä et al, 1998;Kida et al, 1999;Haskell et al, 2000;reviewed in Pearce, 2000). A potentially important additional site of localization has been observed in neurons where CLN3 protein was found in the synaptic region (Järvelä et al, 1999;Haskell et al, 2000;Luiro et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Two Motifs Target Batten Disease Protein CLN3 to Lysosomes in Transfected Nonneuronal and Neuronal Cells

Kyttälä

Ihrke

Vesa

et al. 2004

MBoC

103

145

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Batten disease is a neurodegenerative disorder resulting from mutations in CLN3, a polytopic membrane protein, whose predominant intracellular destination in nonneuronal cells is the lysosome. The topology of CLN3 protein, its lysosomal targeting mechanism, and the development of Batten disease are poorly understood. We provide experimental evidence that both the N and C termini and one large loop domain of CLN3 face the cytoplasm. We have identified two lysosomal targeting motifs that mediate the sorting of CLN3 in transfected nonneuronal and neuronal cells: an unconventional motif in the long C-terminal cytosolic tail consisting of a methionine and a glycine separated by nine amino acids [M(X)9G], and a more conventional dileucine motif, located in the large cytosolic loop domain and preceded by an acidic patch. Each motif on its own was sufficient to mediate lysosomal targeting, but optimal efficiency required both. Interestingly, in primary neurons, CLN3 was prominently seen both in lysosomes in the cell body and in endosomes, containing early endosomal antigen-1 along neuronal processes. Because there are few lysosomes in axons and peripheral parts of dendrites, the presence of CLN3 in endosomes of neurons may be functionally important. Endosomal association of the protein was independent of the two lysosomal targeting motifs.

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Analysis of Intracellular Distribution and Trafficking of the CLN3 Protein in Fusion with the Green Fluorescent Proteinin Vitro

Cited by 39 publications

References 30 publications

The juvenile Batten disease protein, CLN3, and its role in regulating anterograde and retrograde post-Golgi trafficking

The juvenile Batten disease protein, CLN3, and its role in regulating anterograde and retrograde post-Golgi trafficking

Osmotic Stress Changes the Expression and Subcellular Localization of the Batten Disease Protein CLN3

Two Motifs Target Batten Disease Protein CLN3 to Lysosomes in Transfected Nonneuronal and Neuronal Cells

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