# Mammalian fatty acid synthase and O-GlcNAc transferase preferentially interact via their respective N-terminal regions

## TL;DR

This study shows that FASN and OGT, two enzymes linked to cancer, interact mainly through their N-terminal regions, with OGT adding sugar molecules to FASN.

## Contribution

The paper identifies the N-terminal regions as the primary interaction site between FASN and OGT and links stronger interaction to increased glycosylation.

## Key findings

- FASN and OGT preferentially interact via their N-terminal regions.
- Higher OGT interaction with FASN correlates with increased glycosylation of FASN.
- AlphaFold modeling reveals the interaction surface between FASN and OGT.

## Abstract

Fatty Acid Synthase (FASN) is a central enzyme in the de novo lipogenesis pathway. By producing fatty acids, FASN is implicated in numerous crucial cellular processes, but it is also frequently overexpressed in cancer. O-GlcNAc Transferase (OGT) governs the addition of N-acetylglucosamine residues onto cytosolic, nuclear and mitochondrial proteins. Like FASN, OGT actively participates in carcinogenesis. We previously showed that OGT regulates FASN in different ex vivo and in vivo models. Reciprocally, FASN promotes OGT expression and activity. The two enzymes physically interact together and contribute to cancer cell survival. It is therefore fundamental to define the respective interaction region of each enzyme to explore new therapeutic solutions for patients suffering from cancer. By using the hepatocarcinoma cell line Hep3B, we show thanks to two series of deletion mutants that both enzymes preferentially interact via their respective N-terminal regions. Analysis of the O-GlcNAc status of the various FASN deletion mutants shows that stronger interaction with OGT correlates with higher glycosylation, suggesting that OGT catalyzes the transfer of GlcNAc with limited substrate specificity.

•FASN and OGT physically interact together via their respective N-terminal region.•More OGT interacts with a N-terminus deletant version of FASN, more this construct is glycosylated.•Modeling with AlphaFold reveals the interaction surface of both partners.

FASN and OGT physically interact together via their respective N-terminal region.

More OGT interacts with a N-terminus deletant version of FASN, more this construct is glycosylated.

Modeling with AlphaFold reveals the interaction surface of both partners.