Regulation of SESAME-mediated H3T11 phosphorylation by glycolytic enzymes and metabolites

<div><p>Cancer cells prefer aerobic glycolysis, but little is known about the underlying mechanism. Recent studies showed that the rate-limiting glycolytic enzymes, pyruvate kinase M2 (PKM2) directly phosphorylates H3 at threonine 11 (H3T11) to regulate gene expression and cell proliferation, revealing its non-metabolic functions in connecting glycolysis and histone modifications. We have reported that the yeast homolog of PKM2, Pyk1 phosphorylates H3T11 to regulate gene expression and oxidative stress resistance. But how glycolysis regulates H3T11 phosphorylation remains unclear. Here, using a series of glycolytic enzyme mutants and commercial available metabolites, we investigated the role of glycolytic enzymes and metabolites on H3T11 phosphorylation. Mutation of glycolytic genes including phosphoglucose isomerase (<i>PGI1</i>), enolase (<i>ENO2</i>), triosephosphate isomerase (<i>TPI1</i>), or folate biosynthesis enzyme (<i>FOL3</i>) significantly reduced H3T11 phosphorylation. Further study demonstrated that glycolysis regulates H3T11 phosphorylation by fueling the substrate, phosphoenonylpyruvate and the coactivator, FBP to Pyk1. Thus, our results provide a comprehensive view of how glycolysis modulates H3T11 phosphorylation.</p></div

Highly Selective Detection of 5‑Methylcytosine in Genomic DNA Based on Asymmetric PCR and Specific DNA Damaging Reagents

DNA methylation is a significant epigenetic modification of the genome that is involved in regulating many cellular processes. An increasing number of human diseases have been discovered to be associated with aberrant DNA methylation, and aberrant DNA methylation has been deemed to be a potential biomarker for diseases such as cancers. A safe, nontoxic, and sensitive method for accurate detection of 5-methylcytosine in genomic DNA is extremely useful for early diagnosis and therapy of cancers. In this paper, we established a novel system to detect 5-methylcytosine, which is based on bisulfite treatment, asymmetric PCR, and specific DNA damaging reagents. Our method could be used for identifying the loci of 5mC in genomic DNA and detecting the DNA methylation levels in tissues as well

Application of <i>N</i>‑Halogeno‑<i>N</i>‑sodiobenzenesulfonamide Reagents to the Selective Detection of 5‑Methylcytosine in DNA Sequences

To surmount the challenges of the locus determination and accurate quantification of 5-methyl-2′-deoxycytidine (^5MedC) in DNA fragments that contain multiple ^5MedC residues, we designed and synthesized two <i>N</i>-halogeno-<i>N</i>-sodiobenzenesulfonamide reagents that provide a new chemical method for probing ^5MedC in DNA sequences. When the strategy we provided was combined with β-glucosyltransferase, ^5MedC could be distinguished from 5-hydroxymethyl-2′-deoxycytidine (^5hmdC) and deoxycytidine (dC) through the introduction of a glucose moiety to the hydroxyl group of ^5hmdC