Abstract
The sulfation of tyrosine residues is an important post-translational modification involved in the regulation of protein function. We examined the activity of worm tyrosylprotein sulfotransferase (TPST-1) on a typical cuticle collagen, ROL-6, in C. elegans. We verified that TPST-1 sulfates three tyrosine residues of ROL-6 in vitro. We found that these tyrosine residues are important for the secretion of ROL-6::GFP. Mutant ROL-6::GFP proteins that contain more than two substitutions of the target tyrosine residues are severely deficient in cuticle localization. Consistently, knock down of tpst-1 blocked the cuticle localization of ROL-6::GFP. Therefore, the sulfation of ROL-6 by TPST-1 is critical for the proper localization of ROL-6. We also confirmed that worm TPST-1 is localized to the trans-Golgi network (TGN). Our results indicate that TPST-1 regulates cuticle organization by promoting the transport of ROL-6 from the TGN to the cuticle.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Blotting, Western
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Caenorhabditis elegans / enzymology
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Caenorhabditis elegans / genetics
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Caenorhabditis elegans / metabolism*
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Caenorhabditis elegans Proteins / genetics
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Caenorhabditis elegans Proteins / metabolism*
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Cell Line
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Collagen / genetics
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Collagen / metabolism*
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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HeLa Cells
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Humans
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Microscopy, Fluorescence
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Molecular Sequence Data
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Mutation
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Protein Transport
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RNA Interference
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Sulfates / metabolism
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Sulfotransferases / genetics
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Sulfotransferases / metabolism*
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Transfection
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Tyrosine / genetics
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Tyrosine / metabolism
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trans-Golgi Network / metabolism
Substances
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Caenorhabditis elegans Proteins
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Recombinant Fusion Proteins
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Sulfates
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rol-6 protein, C elegans
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Green Fluorescent Proteins
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Tyrosine
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Collagen
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Sulfotransferases
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tpst-1 protein, C elegans