DNA Probe Pulldown Screening Uncovers O-GlcNAcylation Modulation of Transcription Factor DNA Interactions DOI

Guofang Li,

Fanxu Meng, Xiaomin Zhong

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: April 14, 2025

Abstract O-linked β-N-acetylglucosamine (O-GlcNAc), a critical post-translational modification (PTM) predominantly found in the nucleus, plays substantial role regulating gene expression by modulating transcription factors (TFs) activity. However, quantitative analysis investigating influence of O-GlcNAcylation on protein-DNA interactions at proteome scale remains undone. Herein, pulldown screening approach using consensus TF response element (catTFRE) was employed to unravel impact fluctuating levels DNA binding efficiency endogenous TFs/co-factors. Utilizing proteomics, we identified enhancement capacity 241 nuclear proteins (NPs) sequences due elevated O-GlcNAcylation, whereas decrease observed for 2 NPs concurrently. Intriguingly, elevation significantly enhanced 146 TFs/co-factors specific sequences. We further established that several Forkhead family TFs, including FOXA1 and FOXC1, notably enhances their living cells. Our research presents an efficacious assessing between DNA. This our understanding regulation transcription.

Language: Английский

Understanding the redundant functions of the m6A-binding YTHDF proteins DOI Open Access
Sara Zaccara, Samie R. Jaffrey

RNA, Journal Year: 2024, Volume and Issue: 30(5), P. 468 - 481

Published: March 26, 2024

N 6 -methyladenosine (m A) is the most prevalent modified nucleotide in mRNA, and it has important functions mRNA regulation. However, our understanding of specific m A along with its cytosolic readers, YTHDF proteins, changed substantially recent years. The original view was that different sites within an could have depending on which paralog bound to it, YTHDF1 inducing translation, while YTHDF2 induced degradation. As a result, each proposed unique physiologic roles arise from their binding properties regulatory effects mRNA. More data called much this into question, showing all bind proteins equal ability, single primary function three mediate Here, we describe diverse technical concerns led model being questioned newer overturned new function. We also discuss how any remaining questions about can be readily resolved.

Language: Английский

Citations

7
DNA Probe Pulldown Screening Uncovers O-GlcNAcylation Modulation of Transcription Factor DNA Interactions DOI

Guofang Li,

Fanxu Meng, Xiaomin Zhong

et al.

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: April 14, 2025

Abstract O-linked β-N-acetylglucosamine (O-GlcNAc), a critical post-translational modification (PTM) predominantly found in the nucleus, plays substantial role regulating gene expression by modulating transcription factors (TFs) activity. However, quantitative analysis investigating influence of O-GlcNAcylation on protein-DNA interactions at proteome scale remains undone. Herein, pulldown screening approach using consensus TF response element (catTFRE) was employed to unravel impact fluctuating levels DNA binding efficiency endogenous TFs/co-factors. Utilizing proteomics, we identified enhancement capacity 241 nuclear proteins (NPs) sequences due elevated O-GlcNAcylation, whereas decrease observed for 2 NPs concurrently. Intriguingly, elevation significantly enhanced 146 TFs/co-factors specific sequences. We further established that several Forkhead family TFs, including FOXA1 and FOXC1, notably enhances their living cells. Our research presents an efficacious assessing between DNA. This our understanding regulation transcription.

Language: Английский

Citations

0