ST6Gal-I-mediated sialylation of the epidermal growth factor receptor modulates cell mechanics and enhances invasion

• Type of Material: Article
• Authors: Tejeshwar C Rao, University of Alabama Birmingham; Reena R Beggs, University of Alabama Birmingham; Katherine E Ankenbauer, University of Alabama Birmingham; Jihye Hwang, University of Alabama Birmingham; Victor Pui-Yan Ma, Emory University; Khalid Salaita, Emory University; Susan L Bellis, University of Alabama Birmingham; Alexa L Mattheyses, University of Alabama Birmingham
• Language: English
• Date: 2022-04-01
• Publisher: ELSEVIER
• Publication Version: Final Published Version
• Copyright Statement: © 2022 The Authors
• License: Creative Commons Attribution 4.0 International
• Final Published Version (URL): http://dx.doi.org/10.1016/j.jbc.2022.101726
• Title of Journal or Parent Work: JOURNAL OF BIOLOGICAL CHEMISTRY
• Volume: 298
• Issue: 4
• Start Page: 101726
• End Page: 101726
• Grant/Funding Information: This work was supported by funding to A. L. M. from the National Science Foundation (NSF) CAREER (1832100), to A. L. M. and K. S. from the National Institutes of Health (NIH) (R01GM131099), to S. L. B. from the NIH (U01CA233581 and R01CA225177), and to V. P.-Y. M. from the National Cancer Institute (NCI) (K00CA223074). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
• Keywords: PROTECTS TUMOR-CELLS; TYROSINE KINASES; INTEGRIN; Biochemistry & Molecular Biology; SIGNAL-TRANSDUCTION; BETA-GALACTOSIDE ALPHA-2,6-SIALYLTRANSFERASE; Life Sciences & Biomedicine; CROSS-TALK; OVARIAN-CANCER; SIALIC-ACID; Science & Technology; EGFR LIGANDS; N-GLYCANS
• Research Categories: Biology, Cell; Chemistry, General

Abstract

Heterogeneity within the glycocalyx influences cell adhesion mechanics and signaling. However, the role of specific glycosylation subtypes in influencing cell mechanics via alterations of receptor function remains unexplored. It has been shown that the addition of sialic acid to terminal glycans impacts growth, development, and cancer progression. In addition, the sialyltransferase ST6Gal-I promotes epidermal growth factor receptor (EGFR) activity, and we have shown EGFR is an 'allosteric mechano-organizer' of integrin tension. Here, we investigated the impact of ST6Gal-I on cell mechanics. Using DNA-based tension gauge tether probes of variable thresholds, we found that high ST6Gal-I activity promotes increased integrin forces and spreading in Cos-7 and OVCAR3, OVCAR5, and OV4 cancer cells. Further, employing inhibitors and function-blocking antibodies against β1, β3, and β5 integrins and ST6Gal-I targets EGFR, tumor necrosis factor receptor, and Fas cell surface death receptor, we validated that the observed phenotypes are EGFR-specific. We found that while tension, contractility, and adhesion are extracellularsignal- regulated kinase pathway-dependent, spreading, proliferation, and invasion are phosphoinositide 3-kinase-Akt serine/threonine kinase dependent. Using total internal reflection fluorescence microscopy and flow cytometry, we also show that high ST6Gal-I activity leads to sustained EGFR membrane retention, making it a key regulator of cell mechanics. Our findings suggest a novel sialylation-dependent mechanism orchestrating cellular mechanics and enhancing cell motility via EGFR signaling.