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Title: Discovery of an O-mannosylation pathway selectively serving cadherins and protocadherins

Journal Article · · Proceedings of the National Academy of Sciences of the United States of America
 [1];  [1]; ORCiD logo [1];  [1];  [2];  [2];  [2];  [1];  [2];  [2];  [1];  [1]; ORCiD logo [3]
  1. Univ. of Copenhagen (Denmark)
  2. Columbia Univ., New York, NY (United States)
  3. Univ. of Copenhagen (Denmark); Rockefeller Univ., New York, NY (United States)
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The cadherin (cdh) superfamily of adhesion molecules carry O-linked mannose (O-Man) glycans at highly conserved sites localized to specific β-strands of their extracellular cdh (EC) domains. These O-Man glycans do not appear to be elongated like O-Man glycans found on α-dystroglycan (α-DG), and we recently demonstrated that initiation of cdh/protocadherin (pcdh) O-Man glycosylation is not dependent on the evolutionary conserved POMT1/POMT2 enzymes that initiate O-Man glycosylation on α-DG. Here, we used a CRISPR/Cas9 genetic dissection strategy combined with sensitive and quantitative O-Man glycoproteomics to identify a homologous family of four putative protein O-mannosyltransferases encoded by the TMTC1–4 genes, which were found to be imperative for cdh and pcdh O-Man glycosylation. KO of all four TMTC genes in HEK293 cells resulted in specific loss of cdh and pcdh O-Man glycosylation, whereas combined KO of TMTC1 and TMTC3 resulted in selective loss of O-Man glycans on specific β-strands of EC domains, suggesting that each isoenzyme serves a different function. In addition, O-Man glycosylation of IPT/TIG domains of plexins and hepatocyte growth factor receptor was not affected in TMTC KO cells, suggesting the existence of yet another O-Man glycosylation machinery. Furthermore, our study demonstrates that regulation of O-mannosylation in higher eukaryotes is more complex than envisioned, and the discovery of the functions of TMTCs provide insight into cobblestone lissencephaly caused by deficiency in TMTC3.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
Marie Sklodowska-Curie Grant; National Science Foundation (NSF); National Inst. of Health; Danish National Research Foundation
Grant/Contract Number:
704228; MCB-1412472; R01-GM107571; R01GM118584; DNRF107
OSTI ID:
1406612
Journal Information:
Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, Issue 42; ISSN 0027-8424
Publisher:
National Academy of SciencesCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 53 works
Citation information provided by
Web of Science

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Cited By (9)

Incorporation of desmocollin‐2 into the plasma membrane requires N ‐glycosylation at multiple sites journal April 2019
Glycosyltransferase genes that cause monogenic congenital disorders of glycosylation are distinct from glycosyltransferase genes associated with complex diseases journal March 2018
Structures and functions of invertebrate glycosylation journal January 2019
Deletion of Tmtc4 activates the unfolded protein response and causes postnatal hearing loss journal September 2018
Glycosylation in the Tumor Microenvironment: Implications for Tumor Angiogenesis and Metastasis journal June 2019
Membrane Topological Model of Glycosyltransferases of the GT-C Superfamily journal September 2019
Predicted glycosyltransferases promote development and prevent spurious cell clumping in the choanoflagellate S. rosetta journal December 2018
Family-wide Structural and Biophysical Analysis of Binding Interactions among Non-clustered δ-Protocadherins journal February 2020
Monitoring Protein Dynamics in Protein O-Mannosyltransferase Mutants In Vivo by Tandem Fluorescent Protein Timers journal October 2018

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59 BASIC BIOLOGICAL SCIENCES
glycoproteomics
O-glycosylation
glycosyltransferase
mass spectrometry
gene editing