O- Fucosylation of ADAMTSL2 is required for secretion and is impacted by geleophysic dysplasia-causing mutations.

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  • Additional Information
    • Source:
      Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE
    • Publication Information:
      Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
      Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology
    • Subject Terms:
    • Abstract:
      ADAMTSL2 mutations cause an autosomal recessive connective tissue disorder, geleophysic dysplasia 1 (GPHYSD1), which is characterized by short stature, small hands and feet, and cardiac defects. ADAMTSL2 is a matricellular protein previously shown to interact with latent transforming growth factor-β binding protein 1 and influence assembly of fibrillin 1 microfibrils. ADAMTSL2 contains seven thrombospondin type-1 repeats (TSRs), six of which contain the consensus sequence for O -fucosylation by protein O- fucosyltransferase 2 (POFUT2). O- fucose-modified TSRs are subsequently elongated to a glucose β1-3-fucose (GlcFuc) disaccharide by β1,3-glucosyltransferase (B3GLCT). B3GLCT mutations cause Peters Plus Syndrome (PTRPLS), which is characterized by skeletal defects similar to GPHYSD1. Several ADAMTSL2 TSRs also have consensus sequences for C- mannosylation. Six reported GPHYSD1 mutations occur within the TSRs and two lie near O- fucosylation sites. To investigate the effects of TSR glycosylation on ADAMTSL2 function, we used MS to identify glycan modifications at predicted consensus sequences on mouse ADAMTSL2. We found that most TSRs were modified with the GlcFuc disaccharide at high stoichiometry at O -fucosylation sites and variable mannose stoichiometry at C- mannosylation sites. Loss of ADAMTSL2 secretion in POFUT2 -/- but not in B3GLCT -/- cells suggested that impaired ADAMTSL2 secretion is not responsible for skeletal defects in PTRPLS patients. In contrast, secretion was significantly reduced for ADAMTSL2 carrying GPHYSD1 mutations (S641L in TSR3 and G817R in TSR6), and S641L eliminated O- fucosylation of TSR3. These results provide evidence that abnormalities in GPHYSD1 patients with this mutation are caused by loss of O- fucosylation on TSR3 and impaired ADAMTSL2 secretion.
      Competing Interests: Conflict of interest—The authors declare that they have no conflicts of interest with the contents of this article.
      (© 2020 Zhang et al.)
    • References:
      Mol Biol Cell. 1998 Feb;9(2):301-9. (PMID: 9450956)
      Proc Natl Acad Sci U S A. 2017 Mar 7;114(10):2574-2579. (PMID: 28202721)
      J Biol Chem. 2009 Nov 13;284(46):31493-7. (PMID: 19734141)
      FASEB J. 2019 Feb;33(2):2707-2718. (PMID: 30303737)
      J Biol Chem. 2008 Mar 21;283(12):7354-60. (PMID: 18199743)
      Proc Natl Acad Sci U S A. 2005 Apr 19;102(16):5679-84. (PMID: 15824322)
      Arch Biochem Biophys. 1986 Sep;249(2):533-45. (PMID: 2428310)
      J Biol Chem. 2006 Apr 7;281(14):9385-92. (PMID: 16464858)
      Protein Eng. 1990 Apr;3(5):433-42. (PMID: 2349213)
      Cell. 1975 Oct;6(2):121-8. (PMID: 1182798)
      Sci Rep. 2017 Feb 08;7:41871. (PMID: 28176809)
      J Biol Chem. 2001 Mar 2;276(9):6485-98. (PMID: 11067851)
      J Proteome Res. 2011 Dec 2;10(12):5296-301. (PMID: 22010998)
      Clin Genet. 2014 Aug;86(2):142-8. (PMID: 23889335)
      Mol Cell. 2015 May 21;58(4):575-85. (PMID: 26000843)
      Am J Hum Genet. 2006 Sep;79(3):562-6. (PMID: 16909395)
      Glycobiology. 2013 Feb;23(2):188-98. (PMID: 23045360)
      Dev Biol. 2010 Oct 1;346(1):25-38. (PMID: 20637190)
      J Biol Chem. 2006 Apr 7;281(14):9393-9. (PMID: 16464857)
      Sci Rep. 2016 Sep 30;6:33974. (PMID: 27687499)
      Matrix Biol. 2007 Jul;26(6):431-41. (PMID: 17509843)
      Mol Cell. 2013 Apr 25;50(2):295-302. (PMID: 23562325)
      Nat Genet. 2008 Sep;40(9):1119-23. (PMID: 18677313)
      Curr Biol. 2015 Feb 2;25(3):286-295. (PMID: 25544610)
      Hum Mol Genet. 2019 Dec 15;28(24):4053-4066. (PMID: 31600785)
      Mol Genet Metab Rep. 2019 Sep 05;21:100504. (PMID: 31516831)
      Nucleic Acids Res. 2019 Jan 8;47(D1):D442-D450. (PMID: 30395289)
      Methods Enzymol. 2010;480:401-16. (PMID: 20816219)
      Dis Model Mech. 2015 May;8(5):487-99. (PMID: 25762570)
      Cold Spring Harb Perspect Med. 2012 May;2(5):a006627. (PMID: 22553494)
      J Med Genet. 2011 Jun;48(6):417-21. (PMID: 21415077)
      Nature. 2014 Jul 24;511(7510):466-70. (PMID: 24896188)
      Nat Chem Biol. 2016 Apr;12(4):240-6. (PMID: 26854667)
      J Biol Chem. 2007 Jun 8;282(23):17024-31. (PMID: 17395588)
      Curr Opin Struct Biol. 2019 Jun;56:78-86. (PMID: 30690220)
      J Biol Chem. 2007 Jun 8;282(23):17014-23. (PMID: 17395589)
      Elife. 2019 Dec 23;8:. (PMID: 31868591)
      Dev Biol. 2016 Aug 1;416(1):111-122. (PMID: 27297885)
      J Biol Chem. 2009 Oct 30;284(44):30004-15. (PMID: 19671700)
      Matrix Biol. 2015 Sep;47:34-43. (PMID: 25957949)
    • Grant Information:
      R01 CA123071 United States CA NCI NIH HHS; R01 HD090156 United States HD NICHD NIH HHS; R01 HD096030 United States HD NICHD NIH HHS
    • Contributed Indexing:
      Keywords: ADAMTSL2; C-mannosylation; O-fucosylation; TSR; cell biology; extracellular matrix; geleophysic dysplasia; glycoprotein secretion; glycosylation; mass spectrometry
    • Accession Number:
      0 (Adamtsl2 protein, mouse)
      0 (Disaccharides)
      0 (Extracellular Matrix Proteins)
      0 (Recombinant Proteins)
      EC 2.4.- (B3glct protein, mouse)
      EC 2.4.- (Glycosyltransferases)
      EC 2.4.1.- (Fucosyltransferases)
      EC 2.4.1.221 (PoFut2 protein, mouse)
      EC 3.4.24.- (ADAMTS Proteins)
      PHA4727WTP (Mannose)
    • Subject Terms:
      Acromicric dysplasia
    • Publication Date:
      Date Created: 20200911 Date Completed: 20210309 Latest Revision: 20211114
    • Publication Date:
      20231215
    • Accession Number:
      PMC7667967
    • Accession Number:
      10.1074/jbc.RA120.014557
    • Accession Number:
      32913123