Fucosylated Proteome Profiling Identifies a Fucosylated, Non-Ribosomal, Stress-Responsive Species of Ribosomal Protein S3

Abstract

Alterations in genes encoding for proteins that control fucosylation are known to play causative roles in several developmental disorders, such as Dowling-Degos disease 2 and congenital disorder of glycosylation type IIc (CDGIIc). Recent studies have provided evidence that changes in fucosylation can contribute to the development and progression of several different types of cancers. It is therefore important to gain a detailed understanding of how fucosylation is altered in disease states so that interventions may be developed for therapeutic purposes. In this report, we find that fucosylation occurs on many intracellular proteins. This is an interesting finding, as the fucosylation machinery is restricted to the secretory pathway and is thought to predominately affect cell-membrane-bound and secreted proteins. We find that Ribosomal protein S3 (RPS3) is fucosylated in normal tissues and in cancer cells, and that the extent of its fucosylation appears to respond to stress, including MAPK inhibitors, suggesting a new role in posttranslational protein function. Our data identify a new ribosome-independent species of fucosylated RPS3 that interacts with proteins involved in posttranscriptional regulation of RNA, such as Heterogeneous nuclear ribonucleoprotein U (HNRNPU), as well as with a predominance of non-coding RNAs. These data highlight a novel role for RPS3, which, given previously reported oncogenic roles for RPS3, might represent functions that are perturbed in pathologies such as cancer. Together, our findings suggest a previously unrecognized role for fucosylation in directly influencing intracellular protein functions.

Androgen drives melanoma invasiveness and metastatic spread by inducing tumorigenic fucosylation

“…5c ). Lectin-mediated PLA (LPLA, a method developed in our laboratory 15 , 16 that allows for the visualization of specific fucosylated proteins) also confirmed that knockdown or overexpression of FUT4 resulted in reduced or increased, respectively, cellular levels of fucosylated L1CAM (fuco-L1CAM) (Fig. 5c and Supplementary Fig.…”

“…Indeed, in cancer, aberrant levels of fucosylated proteins have been reported 10 , but their specific mechanistic roles in cancer pathogenesis are unclear. Aberrant structural subtypes of fucosylation ( e.g ., core fucosylation) have been reported in melanoma 12 , 13 , and we previously discovered that global fucosylation is deregulated during melanoma progression, impacting motility and RNA processing of melanoma cells 14 , 15 and altering crucial tumor:immunological interactions that are required for anti-tumor immunity 16 . Intriguingly, our recent work uncovered that global tumoral fucosylation is significantly lower in male compared to female melanoma patients, suggesting sex-associated divergence in fucosylation-regulated melanoma biology 16 .…”

High-Throughput Mass Spectrometry Analysis of N-Glycans and Protein Markers after FUT8 Knockdown in the Syngeneic SW480/SW620 Colorectal Cancer Cell Model

“…Proteins related to the damage response deserve attention as they may be related to radio- and/or chemoresistance or drug sensitivity, thus encompassing potential drug targets through the inhibition of FucT-8 activity (cluster c, Figure ). In this sense, it has recently been described that deregulated fucosylation can affect many intracellular proteins, including the ribosomal protein S3, which participates in nuclear DNA repair . Indeed, the fucosylation and sialylation inhibitor pictilisib affected the DNA repair capacity of lung cancer cells .…”

“…In this sense, it has recently been described that deregulated fucosylation can affect many intracellular proteins, including the ribosomal protein S3, which participates in nuclear DNA repair. 113 Indeed, the fucosylation and sialylation inhibitor pictilisib affected the DNA repair capacity of lung cancer cells. 114 The histone H2AFJ (cluster e) is usually expressed in luminal epithelial cells, although its role is still poorly understood.…”

Ribosome-Directed Therapies in Cancer

“…It is thought that fucose binding lectin and many other intracellular proteins undergoes post-translation fucosylation. The ribosomal protein S3 (RPS3) is fucosylated in human cancer cells and normal mouse tissue [ 199 ].…”