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

Ida Signe Bohse Larsen; Yoshiki Narimatsu; Hiren Jitendra Joshi; Lina Siukstaite; Oliver J Harrison; Julia Brasch; Kerry M Goodman; Lars Hansen; Lawrence Shapiro; Barry Honig; Sergey Y Vakhrushev; Henrik Clausen; Adnan Halim

doi:10.1073/pnas.1708319114

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

Ida Signe Bohse Larsen, Yoshiki Narimatsu, Hiren Jitendra Joshi, Lina Siukstaite, Oliver J Harrison, Julia Brasch, Kerry M Goodman, Lars Hansen, Lawrence Shapiro, Barry Honig, Sergey Y Vakhrushev, Henrik Clausen, Adnan Halim

Glycomics Program

35 Citations (Scopus)

Abstract

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. 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.

Original language	English
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	114
Issue number	42
Pages (from-to)	11163-11168
Number of pages	6
ISSN	0027-8424
DOIs	https://doi.org/10.1073/pnas.1708319114
Publication status	Published - 17 Oct 2017

Keywords

Journal Article

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Larsen, I. S. B., Narimatsu, Y., Joshi, H. J., Siukstaite, L., Harrison, O. J., Brasch, J., Goodman, K. M., Hansen, L., Shapiro, L., Honig, B., Vakhrushev, S. Y., Clausen, H., & Halim, A. (2017). Discovery of an O-mannosylation pathway selectively serving cadherins and protocadherins. Proceedings of the National Academy of Sciences of the United States of America, 114(42), 11163-11168. https://doi.org/10.1073/pnas.1708319114

Discovery of an O-mannosylation pathway selectively serving cadherins and protocadherins. / Larsen, Ida Signe Bohse ; Narimatsu, Yoshiki ; Joshi, Hiren Jitendra et al.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 42, 17.10.2017, p. 11163-11168.

Research output: Contribution to journal › Journal article › Research › peer-review

Larsen, ISB , Narimatsu, Y , Joshi, HJ, Siukstaite, L, Harrison, OJ, Brasch, J, Goodman, KM, Hansen, L, Shapiro, L, Honig, B, Vakhrushev, SY , Clausen, H & Halim, A 2017, 'Discovery of an O-mannosylation pathway selectively serving cadherins and protocadherins', Proceedings of the National Academy of Sciences of the United States of America, vol. 114, no. 42, pp. 11163-11168. https://doi.org/10.1073/pnas.1708319114

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AU - Larsen, Ida Signe Bohse

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AU - Joshi, Hiren Jitendra

AU - Siukstaite, Lina

AU - Harrison, Oliver J

AU - Brasch, Julia

AU - Goodman, Kerry M

AU - Hansen, Lars

AU - Shapiro, Lawrence

AU - Honig, Barry

AU - Vakhrushev, Sergey Y

AU - Clausen, Henrik

AU - Halim, Adnan

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AB - 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. 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.

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JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 42

ER -

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

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