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GalNAc-T4 putatively modulates the estrogen regulatory network through FOXA1 glycosylation in human breast cancer cells

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Abstract

GALNT4 belongs to a family of N-acetylgalactosaminyltransferases, which catalyze the transfer of GalNAc to Serine or Threonine residues in the initial step of mucin-type O-linked protein glycosylation. This glycosylation type is the most complex post-translational modification of proteins, playing important roles during cellular differentiation and in pathological disorders. Most of the breast cancer subtypes are estrogen receptor positive, and hence, the estrogen pathway represents a key regulatory network. We investigated the expression of GalNAc-T4 in a panel of mammary epithelial cell lines and found its expression is associated with the estrogen status of the cells. FOXA1, a key transcription factor, functions to promote estrogen responsive gene expression by acting as a cofactor to estrogen receptor alpha (ERα), but all the aspects of this regulatory mechanism are not fully explored. This study found that knockdown of GALNT4 expression in human breast cancer cells attenuated the protein expression of ERα, FOXA1, and Cyclin D1. Further, our immunoprecipitation assays depicted the possibility of FOXA1 to undergo O-GalNAc modifications with a decrease of GalNAc residues in the GALNT4 knockdown cells and also impairment in the FOXA1–ERα association. Rescuing GALNT4 expression could restore the interaction as well as the glycosylation of FOXA1. Together, these findings suggest a key role for GalNAc-T4 in the estrogen pathway through FOXA1 glycosylation.

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Abbreviations

GALNT4 :

Polypeptide N-acetylgalactosaminyltransferase 4

ERα:

Estrogen receptor alpha

ERβ:

Estrogen receptor beta

FOXA1:

Forkhead box protein A1

VVL:

Vicia Villosa Lectin

UPR:

Unfolded protein response

ERAD:

Endoplasmic reticulum-associated degradation

CRD:

Carbohydrate recognition domain

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Acknowledgments

This study was supported by grants from the National Program on Key Basic Research Project (973 Program) (No. 2012CB822103), the National Natural Science Foundation of China (No. 31170774, 31570802), the Fundamental Research Funds for the Central Universities and Development Fund for Collaborative Innovation Center of Glycoscience of Shandong University. We thank Professors Baiqu Huang (Northeast Normal University Changchun, China) and Eric Paul Bennett (Copenhagen center for Glycomics, Denmark) for their collaboration.

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Authors and Affiliations

  1. Department of Biochemistry, Institute of Glycobiology, Dalian Medical University, 9 South Lvshun Road Western Section, Dalian, 116044, China

    Bachir Niang, Liyuan Jin, Xixi Chen, Xiaohan Guo, Hongshuo Zhang, Arshad Ahmed Padhiar & Jianing Zhang

  2. School of Life Science and Medicine, Dalian University of Technology, 2 Linggong Road, Dalian, 116024, China

    Qiong Wu & Jianing Zhang

  3. National Glycoengineering Research Center and State Key Laboratory of Microbial Technology, Shandong University, Jinan, 250100, China

    Min Xiao

  4. Department of Pathology, Northwestern University Feinberg School of Medicine, 303 E. Chicago Ave, Chicago, IL, 60611, USA

    Deyu Fang

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  1. Bachir Niang
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  2. Liyuan Jin
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Correspondence to Jianing Zhang.

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Supplementary material 1

Human glycosylation array. The clustergram represents fold-regulation of 84 glycosyltransferases in three mammary epithelial cells MCF10A, T47D and MDAMB231, including GALNT4 which is the first glycosyltransferase on the top of this representation. Note that MCF10A was considered as control. Supplementary material 1 (TIFF 9311 kb)

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Niang, B., Jin, L., Chen, X. et al. GalNAc-T4 putatively modulates the estrogen regulatory network through FOXA1 glycosylation in human breast cancer cells. Mol Cell Biochem 411, 393–402 (2016). https://doi.org/10.1007/s11010-015-2601-1

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  • DOI: https://doi.org/10.1007/s11010-015-2601-1

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