# African Swine Fever Virus R238L and R298L Disrupt Lung Cell Collagen Formation and Cell Adhesion Pathway by Targeting Transcription Factors Containing zf-C2H2 Domain

## TL;DR

This study shows how two proteins from the African Swine Fever Virus disrupt lung cell structure by targeting specific host transcription factors.

## Contribution

The novel contribution is identifying how ASFV proteins R238L and R298L bind to SP2 and KLF6 to disrupt collagen and cell adhesion pathways.

## Key findings

- ASFV proteins R238L and R298L bind to SP2 and KLF6 transcription factors.
- Binding reduces transcription of genes involved in collagen formation and cell adhesion.
- This disruption may aid viral pathogenesis by weakening lung cell structure.

## Abstract

Collagen formation (CF) and cell adhesion (CH) are essential biological processes that maintain cellular structure and communication. This study aimed to investigate whether and how the African Swine Fever Virus (ASFV) affects these host pathways. We identified two host transcription factors, SP2 and KLF6, which regulate the transcription of CF and CH genes. Furthermore, we found that two viral proteins, R238L and R298L, can bind directly to SP2 and KLF6 and reduce the transcription of their target genes.

The regulatory mechanisms of collagen formation and cell adhesion pathways during African Swine Fever Virus (ASFV) infection remain poorly understood. This study aims to investigate whether ASFV manipulates these pathways by targeting host transcriptional regulators. Through weighted Kendall correlation analysis of transcription factor binding sites (TFBSs) in differentially expressed genes (DEGs) from the lung tissue of ASFV-recovered pigs, we identified SP2 and KLF6 as key transcription factors (TFs) associated with collagen synthesis and cell adhesion, respectively. Domain–domain interaction prediction, followed by Förster resonance energy transfer (FRET) assays, confirmed that the ASFV proteins R238L and R298L directly bind to the zf-C2H2 domains of SP2 and KLF6. Furthermore, overexpression of R238L and R298L in HeLa and 3D4/21 cells significantly downregulated SP2 and KLF6 target genes involved in these pathways. Our findings reveal a novel mechanism by which ASFV proteins R238L and R298L interfere with host transcription factors SP2 and KLF6, potentially disrupting collagen matrix integrity and cell adhesion to facilitate viral pathogenesis.