RESUMEN
We herein employ in situ Hi-C with an auxin-inducible degron (AID) system to examine the effect of chromatin remodeling on 3D genome organization in yeast. Eight selected ATP-dependent chromatin remodelers representing various subfamilies contribute to 3D genome organization differently. Among the studied remodelers, the temporary depletions of Chd1p, Swr1p, and Sth1p (a catalytic subunit of the Remodeling the Structure of Chromatin [RSC] complex) cause the most significant defects in intra-chromosomal contacts, and the regulatory roles of these three remodelers in 3D genome organization differ depending on the chromosomal context and cell cycle stage. Furthermore, even though Chd1p and Isw1p are known to share functional similarities/redundancies, their depletions lead to distinct effects on 3D structures. The RSC and cohesin complexes also differentially modulate 3D genome organization within chromosome arm regions, whereas RSC appears to support the function of cohesin in centromeric clustering at G2 phase. Our work suggests that the ATP-dependent chromatin remodelers control the 3D genome organization of yeast through their chromatin-remodeling activities.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Ensamble y Desensamble de Cromatina , Proteínas Cromosómicas no Histona/metabolismo , Nucleosomas/metabolismo , Saccharomycetales/metabolismo , Ciclo Celular/fisiología , Saccharomycetales/genética , CohesinasRESUMEN
It is important to inactivate spore-forming bacteria in foods because their spores are highly resistant to various stresses. Although thermal treatment is an effective inactivation method, the associated high temperatures can cause changes in food quality. Intense pulsed light (IPL) is a nonthermal technique that can effectively improve food safety. This study evaluated the inactivation effects of IPL at various fluences on Bacillus subtilis spores. IPL treatment at a total fluence of 7.40â¯J/cm2 resulted in a 7 log reduction, indicating the potential of IPL to effectively inactivate bacterial spores. The sensitivity of B. subtilis spores to IPL during germination and outgrowth was also measured. The resistance to the IPL increased temporarily until 1â¯h after the start of incubation, and then gradually decreased for longer incubation periods. This temporary increase in resistance at the early stage of incubation was attributed to the leakage of dipicolinic acid from the spores. The results also showed that the inactivation efficiency increases after 1â¯h pre-incubation because the numbers of vegetative cells increased with the incubation time.
Asunto(s)
Bacillus subtilis , Microbiología de Alimentos/métodos , Luz , Esporas Bacterianas/fisiología , Esporas Bacterianas/efectos de la radiación , Bacillus subtilis/fisiología , Bacillus subtilis/efectos de la radiación , Conservación de Alimentos , Viabilidad Microbiana/efectos de la radiación , Ácidos Picolínicos/metabolismo , Esporas Bacterianas/crecimiento & desarrollo , Esporas Bacterianas/metabolismoRESUMEN
In Saccharomyces cerevisiae, the ATP-dependent chromatin remodeler, Chd1p, globally affects nucleosome positioning at coding regions, where nucleosomes are specifically and directionally aligned with respect to the transcription start site (TSS). Various auxiliary domains of remodelers play critical roles by performing specialized functions that are unique to the type of remodeler. Here, we report that yeast Chd1p directly binds to acetylated histone H3K36 (H3K36Ac) via its chromodomain, and that H3K36Ac stimulates the nucleosome sliding activity of Chd1p in vitro. Furthermore, we use genome-wide analysis to demonstrate that H3K36Ac promotes the remodeling activity of Chd1p to maintain chromatin stability at the 5' ends of genes in vivo. Our work linking Chd1p with H3K36Ac provides novel insights into how the nucleosome remodeling activity of Chd1p is controlled near the TSS.
Asunto(s)
Proteínas de Unión al ADN/metabolismo , Histonas/metabolismo , Nucleosomas/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Sitio de Iniciación de la Transcripción , Transcripción Genética/genética , Proteínas de Unión al ADN/genética , Saccharomyces cerevisiae/química , Proteínas de Saccharomyces cerevisiae/genéticaRESUMEN
Dot1 (disruptor of telomeric silencing-1, DOT1L in humans) is the only known enzyme responsible for histone H3 lysine 79 methylation (H3K79me) and is evolutionarily conserved in most eukaryotes. Yeast Dot1p lacks a SET domain and does not methylate free histones and thus may have different actions with respect to other histone methyltransferases. Here we show that Dot1p displays histone chaperone activity and regulates nucleosome dynamics via histone exchange in yeast. We show that a methylation-independent function of Dot1p is required for the cryptic transcription within transcribed regions seen following disruption of the Set2-Rpd3S pathway. Dot1p can assemble core histones to nucleosomes and facilitate ATP-dependent chromatin-remodeling activity through its nucleosome-binding domain, in vitro. Global analysis indicates that Dot1p appears to be particularly important for histone exchange and chromatin accessibility on the transcribed regions of long-length genes. Our findings collectively suggest that Dot1p-mediated histone chaperone activity controls nucleosome dynamics in transcribed regions.