Tyrosylprotein sulfotransferase regulates collagen secretion in Caenorhabditis elegans.
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.
Kim
Tai Hoon
TH
Department of Biology, Yonsei University, Seoul, 120-749, Korea.
Kim
Do Hyun
DH
Nam
Hyung Wook
HW
Park
Sang Yoon
SY
Shim
Jaegal
J
Cho
Jin Won
JW
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D017173:Caenorhabditis elegans / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009154:Mutation; D021381:Protein Transport; D034622:RNA Interference; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013431:Sulfates / Q000378:metabolism; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism*; D014162:Transfection; D014443:Tyrosine / Q000235:genetics / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism
D029742:Caenorhabditis elegans Proteins; D011993:Recombinant Fusion Proteins; D013431:Sulfates; C098282:rol-6 protein, C elegans; D049452:Green Fluorescent Proteins; D014443:Tyrosine; D003094:Collagen; D015238:Sulfotransferases; C512780:tpst-1 protein, C elegans