Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 120
Filtrar
1.
bioRxiv ; 2024 Sep 16.
Artículo en Inglés | MEDLINE | ID: mdl-39345615

RESUMEN

H3K9me3-heterochromatin, established by lysine methyltransferases (KMTs) and compacted by HP1 isoforms, represses alternative lineage genes and DNA repeats. Our understanding of H3K9me3-heterochromatin stability is presently limited to individual domains and DNA repeats. We engineered Suv39h2 KO mouse embryonic stem cells to degrade remaining two H3K9me3-KMTs within one hour and found that both passive dilution and active removal contribute to H3K9me3 decay within 12-24 hours. We discovered four different H3K9me3 decay rates across the genome and chromatin features and transcription factor binding patterns that predict the stability classes. A "binary switch" governs heterochromatin compaction, with HP1 rapidly dissociating from heterochromatin upon KMTs' depletion and a particular threshold level of HP1 limiting pioneer factor binding, chromatin opening, and exit from pluripotency within 12 hr. Unexpectedly, receding H3K9me3 domains unearth residual HP1ß peaks enriched with heterochromatin-inducing proteins. Our findings reveal distinct H3K9me3-heterochromatin maintenance dynamics governing gene networks and repeats that together safeguard pluripotency.

2.
Mol Cell ; 84(16): 3061-3079.e10, 2024 Aug 22.
Artículo en Inglés | MEDLINE | ID: mdl-39121853

RESUMEN

Mouse FOXA1 and GATA4 are prototypes of pioneer factors, initiating liver cell development by binding to the N1 nucleosome in the enhancer of the ALB1 gene. Using cryoelectron microscopy (cryo-EM), we determined the structures of the free N1 nucleosome and its complexes with FOXA1 and GATA4, both individually and in combination. We found that the DNA-binding domains of FOXA1 and GATA4 mainly recognize the linker DNA and an internal site in the nucleosome, respectively, whereas their intrinsically disordered regions interact with the acidic patch on histone H2A-H2B. FOXA1 efficiently enhances GATA4 binding by repositioning the N1 nucleosome. In vivo DNA editing and bioinformatics analyses suggest that the co-binding mode of FOXA1 and GATA4 plays important roles in regulating genes involved in liver cell functions. Our results reveal the mechanism whereby FOXA1 and GATA4 cooperatively bind to the nucleosome through nucleosome repositioning, opening chromatin by bending linker DNA and obstructing nucleosome packing.


Asunto(s)
Microscopía por Crioelectrón , Factor de Transcripción GATA4 , Factor Nuclear 3-alfa del Hepatocito , Nucleosomas , Unión Proteica , Factor Nuclear 3-alfa del Hepatocito/metabolismo , Factor Nuclear 3-alfa del Hepatocito/genética , Nucleosomas/metabolismo , Nucleosomas/genética , Nucleosomas/ultraestructura , Animales , Factor de Transcripción GATA4/metabolismo , Factor de Transcripción GATA4/genética , Factor de Transcripción GATA4/química , Ratones , Cromatina/metabolismo , Cromatina/genética , Histonas/metabolismo , Histonas/genética , Histonas/química , Sitios de Unión , ADN/metabolismo , ADN/genética , ADN/química , Ensamble y Desensamble de Cromatina , Humanos
3.
Bioessays ; 46(10): e2400138, 2024 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-39058903

RESUMEN

Pioneer transcription factors, by virtue of their ability to target nucleosomal DNA in silent chromatin, play crucial roles in eliciting cell fate decisions during development and cellular reprogramming. In addition to their well-established role in chromatin opening to activate gene expression programs, recent studies have demonstrated that pioneer factors have the complementary function of being able to silence the starting cell identity through targeted chromatin repression. Given recent discoveries regarding the repressive aspect of pioneer function, we discuss the basis by which pioneer factors can suppress alternative lineage programs in the context of cell fate control.


Asunto(s)
Cromatina , Factores de Transcripción , Animales , Humanos , Diferenciación Celular/genética , Linaje de la Célula/genética , Cromatina/metabolismo , Cromatina/genética , Nucleosomas/metabolismo , Factores de Transcripción/metabolismo , Factores de Transcripción/genética
4.
bioRxiv ; 2024 Jun 06.
Artículo en Inglés | MEDLINE | ID: mdl-38895478

RESUMEN

During development, H3K9me3 heterochromatin is dynamically rearranged, silencing repeat elements and protein coding genes to restrict cell identity. Enhancer of Rudimentary Homolog (ERH) is an evolutionarily conserved protein originally characterized in fission yeast and recently shown to be required for H3K9me3 maintenance in human fibroblasts, but its function during development remains unknown. Here, we show that ERH is required for proper segregation of the inner cell mass and trophectoderm cell lineages during mouse development by repressing totipotent and alternative lineage programs. During human embryonic stem cell (hESC) differentiation into germ layer lineages, ERH is crucial for silencing naïve and pluripotency genes, transposable elements, and alternative lineage genes. Strikingly, ERH depletion in somatic cells reverts the H3K9me3 landscape to an hESC state and enables naïve and pluripotency gene and transposable element activation during iPSC reprogramming. Our findings reveal a role for ERH in initiation and maintenance of developmentally established gene repression.

5.
Nat Commun ; 15(1): 1761, 2024 Feb 26.
Artículo en Inglés | MEDLINE | ID: mdl-38409161

RESUMEN

Tissue damage elicits cell fate switching through a process called metaplasia, but how the starting cell fate is silenced and the new cell fate is activated has not been investigated in animals. In cell culture, pioneer transcription factors mediate "reprogramming" by opening new chromatin sites for expression that can attract transcription factors from the starting cell's enhancers. Here we report that SOX4 is sufficient to initiate hepatobiliary metaplasia in the adult mouse liver, closely mimicking metaplasia initiated by toxic damage to the liver. In lineage-traced cells, we assessed the timing of SOX4-mediated opening of enhancer chromatin versus enhancer decommissioning. Initially, SOX4 directly binds to and closes hepatocyte regulatory sequences via an overlapping motif with HNF4A, a hepatocyte master regulatory transcription factor. Subsequently, SOX4 exerts pioneer factor activity to open biliary regulatory sequences. The results delineate a hierarchy by which gene networks become reprogrammed under physiological conditions, providing deeper insight into the basis for cell fate transitions in animals.


Asunto(s)
Reprogramación Celular , Cromatina , Animales , Ratones , Diferenciación Celular/genética , Reprogramación Celular/genética , Metaplasia , Factores de Transcripción/metabolismo
6.
Nat Aging ; 4(2): 185-197, 2024 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-38267705

RESUMEN

Sterile inflammation, also known as 'inflammaging', is a hallmark of tissue aging. Cellular senescence contributes to tissue aging, in part, through the secretion of proinflammatory factors collectively known as the senescence-associated secretory phenotype (SASP). The genetic variability of thioredoxin reductase 1 (TXNRD1) is associated with aging and age-associated phenotypes such as late-life survival, activity of daily living and physical performance in old age. TXNRD1's role in regulating tissue aging has been attributed to its enzymatic role in cellular redox regulation. Here, we show that TXNRD1 drives the SASP and inflammaging through the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) innate immune response pathway independently of its enzymatic activity. TXNRD1 localizes to cytoplasmic chromatin fragments and interacts with cGAS in a senescence-status-dependent manner, which is necessary for the SASP. TXNRD1 enhances the enzymatic activity of cGAS. TXNRD1 is required for both the tumor-promoting and immune surveillance functions of senescent cells, which are mediated by the SASP in vivo in mouse models. Treatment of aged mice with a TXNRD1 inhibitor that disrupts its interaction with cGAS, but not with an inhibitor of its enzymatic activity alone, downregulated markers of inflammaging in several tissues. In summary, our results show that TXNRD1 promotes the SASP through the innate immune response, with implications for inflammaging. This suggests that the TXNRD1-cGAS interaction is a relevant target for selectively suppressing inflammaging.


Asunto(s)
Transducción de Señal , Tiorredoxina Reductasa 1 , Animales , Ratones , Senescencia Celular/genética , Inmunidad Innata/genética , Inflamación/genética , Nucleotidiltransferasas/genética , Tiorredoxina Reductasa 1/metabolismo
7.
Trends Genet ; 40(2): 134-148, 2024 02.
Artículo en Inglés | MEDLINE | ID: mdl-37940484

RESUMEN

Pioneer factors are a subclass of transcription factors that can bind and initiate opening of silent chromatin regions. Pioneer factors subsequently regulate lineage-specific genes and enhancers and, thus, activate the zygotic genome after fertilization, guide cell fate transitions during development, and promote various forms of human cancers. As such, pioneer factors are useful in directed cell reprogramming. In this review, we define the structural and functional characteristics of pioneer factors, how they bind and initiate opening of closed chromatin regions, and the consequences for chromatin dynamics and gene expression during cell differentiation. We also discuss emerging mechanisms that modulate pioneer factors during development.


Asunto(s)
Cromatina , Factores de Transcripción , Humanos , Factores de Transcripción/genética , Diferenciación Celular/genética , Reprogramación Celular , Cigoto
9.
Cell Rep ; 42(7): 112748, 2023 07 25.
Artículo en Inglés | MEDLINE | ID: mdl-37405916

RESUMEN

Pioneer transcription factors interact with nucleosomes to scan silent, compact chromatin, enabling cooperative events that modulate gene activity. While at a subset of sites pioneer factors access chromatin by assisted loading with other transcription factors, the nucleosome-binding properties of pioneer factors enable them to initiate zygotic genome activation, embryonic development, and cellular reprogramming. To better understand nucleosome targeting in vivo, we assess whether pioneer factors FoxA1 and Sox2 target stable or unstable nucleosomes and find that they target DNase-resistant, stable nucleosomes, whereas HNF4A, a non-nucleosome binding factor, targets open, DNase-sensitive chromatin. Despite FOXA1 and SOX2 targeting similar proportions of DNase-resistant chromatin, using single-molecule tracking, we find that FOXA1 uses lower nucleoplasmic diffusion and longer residence times while SOX2 uses higher nucleoplasmic diffusion and shorter residence times to scan compact chromatin, while HNF4 scans compact chromatin much less efficiently. Thus, pioneer factors target compact chromatin through distinct processes.


Asunto(s)
Cromatina , Factor Nuclear 3-alfa del Hepatocito , Nucleosomas , Factores de Transcripción SOXB1 , Desoxirribonucleasas/metabolismo , Unión Proteica , Factores de Transcripción/metabolismo , Factor Nuclear 3-alfa del Hepatocito/metabolismo , Factores de Transcripción SOXB1/metabolismo
10.
Science ; 381(6653): 92-100, 2023 07 07.
Artículo en Inglés | MEDLINE | ID: mdl-37410825

RESUMEN

Nanoscale chromatin organization regulates gene expression. Although chromatin is notably reprogrammed during zygotic genome activation (ZGA), the organization of chromatin regulatory factors during this universal process remains unclear. In this work, we developed chromatin expansion microscopy (ChromExM) to visualize chromatin, transcription, and transcription factors in vivo. ChromExM of embryos during ZGA revealed how the pioneer factor Nanog interacts with nucleosomes and RNA polymerase II (Pol II), providing direct visualization of transcriptional elongation as string-like nanostructures. Blocking elongation led to more Pol II particles clustered around Nanog, with Pol II stalled at promoters and Nanog-bound enhancers. This led to a new model termed "kiss and kick", in which enhancer-promoter contacts are transient and released by transcriptional elongation. Our results demonstrate that ChromExM is broadly applicable to study nanoscale nuclear organization.


Asunto(s)
Cromatina , Microscopía Fluorescente , Transcripción Genética , Cigoto , Cromatina/química , Nucleosomas/química , ARN Polimerasa II/química , ARN Polimerasa II/metabolismo , Microscopía Fluorescente/métodos , Animales , Pez Cebra , Embrión no Mamífero , Cigoto/metabolismo , Proteína Homeótica Nanog/química , Proteína Homeótica Nanog/metabolismo
11.
Genes Dev ; 37(1-2): 63-68, 2023 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-37061958
13.
Trends Biochem Sci ; 48(6): 513-526, 2023 06.
Artículo en Inglés | MEDLINE | ID: mdl-36990958

RESUMEN

Heterochromatin is defined as a chromosomal domain harboring repressive H3K9me2/3 or H3K27me3 histone modifications and relevant factors that physically compact the chromatin. Heterochromatin can restrict where transcription factors bind, providing a barrier to gene activation and changes in cell identity. While heterochromatin thus helps maintain cell differentiation, it presents a barrier to overcome during efforts to reprogram cells for biomedical purposes. Recent findings have revealed complexity in the composition and regulation of heterochromatin, and shown that transiently disrupting the machinery of heterochromatin can enhance reprogramming. Here, we discuss how heterochromatin is established and maintained during development, and how our growing understanding of the mechanisms regulating H3K9me3 heterochromatin can be leveraged to improve our ability to direct changes in cell identity.


Asunto(s)
Heterocromatina , Histonas , Histonas/metabolismo , Cromatina , Diferenciación Celular , Factores de Transcripción/metabolismo
14.
bioRxiv ; 2023 Feb 14.
Artículo en Inglés | MEDLINE | ID: mdl-36824858

RESUMEN

Tissue damage elicits cell fate switching through a process called metaplasia, but how the starting cell fate is silenced and the new cell fate is activated has not been investigated in animals. In cell culture, pioneer transcription factors mediate "reprogramming" by opening new chromatin sites for expression that can attract transcription factors from the starting cell's enhancers. Here we report that Sox4 is sufficient to initiate hepatobiliary metaplasia in the adult liver. In lineage-traced cells, we assessed the timing of Sox4-mediated opening of enhancer chromatin versus enhancer decommissioning. Initially, Sox4 directly binds to and closes hepatocyte regulatory sequences via a motif it overlaps with Hnf4a, a hepatocyte master regulator. Subsequently, Sox4 exerts pioneer factor activity to open biliary regulatory sequences. The results delineate a hierarchy by which gene networks become reprogrammed under physiological conditions, providing deeper insight into the basis for cell fate transitions in animals.

15.
Dev Cell ; 58(1): 1-2, 2023 01 09.
Artículo en Inglés | MEDLINE | ID: mdl-36626868

RESUMEN

To induce cell fate changes, do transcription factors engage open domains of chromatin or elicit chromatin opening in a pioneering fashion? In this issue of Developmental Cell, Delás et al. show that the same sonic hedgehog (Shh) inducing signal can yield different neural tube fates by either modality.


Asunto(s)
Proteínas Hedgehog , Factores de Transcripción , Proteínas Hedgehog/metabolismo , Diferenciación Celular , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Tubo Neural/metabolismo , Cromatina , Regulación del Desarrollo de la Expresión Génica
16.
Nat Struct Mol Biol ; 30(1): 31-37, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36536103

RESUMEN

To determine how different pioneer transcription factors form a targeted, accessible nucleosome within compacted chromatin and collaborate with an ATP-dependent chromatin remodeler, we generated nucleosome arrays in vitro with a central nucleosome containing binding sites for the hematopoietic E-Twenty Six (ETS) factor PU.1 and Basic Leucine Zipper (bZIP) factors C/EBPα and C/EBPß. Our long-read sequencing reveals that each factor can expose a targeted nucleosome on linker histone-compacted arrays, but with different nuclease sensitivity patterns. The DNA binding domain of PU.1 binds mononucleosomes, but requires an additional intrinsically disordered domain to bind and open compacted chromatin. The canonical mammalian SWI/SNF (cBAF) remodeler was unable to act upon two forms of locally open chromatin unless cBAF was enabled by a separate transactivation domain of PU.1. cBAF potentiates the PU.1 DNA binding domain to weakly open chromatin in the absence of the PU.1 disordered domain. Our findings reveal a hierarchy by which chromatin is opened and show that pioneer factors can provide specificity for action by nucleosome remodelers.


Asunto(s)
Cromatina , Nucleosomas , Animales , Factores de Transcripción/metabolismo , ADN , Adenosina Trifosfato/metabolismo , Ensamble y Desensamble de Cromatina , Mamíferos/genética
18.
Curr Opin Struct Biol ; 75: 102425, 2022 08.
Artículo en Inglés | MEDLINE | ID: mdl-35863165

RESUMEN

Pioneer transcription factors are able to bind a partially exposed motif on the surface of a nucleosome, enabling the proteins to target sites in silent regions of chromatin that have been compacted by linker histone. The targeting of nucleosomal DNA by pioneer factors has been observed in vitro and in vivo, where binding can promote local nucleosome exposure that allows other transcription factors, nucleosome remodelers, and histone modifiers to engage the chromatin and elicit gene activation or further repression. Pioneer factors thereby establish new gene expression programs during cell fate changes that occur during embryonic development, regeneration, and cancer. Here, we review recent biophysical studies that reveal the structural features and strategies used by pioneer factors to accomplish nucleosome binding and the consequential changes to nucleosomes that can lead to DNA accessibility.


Asunto(s)
Histonas , Nucleosomas , Cromatina , ADN/química , Histonas/metabolismo , Factores de Transcripción/metabolismo
19.
Nat Cell Biol ; 24(5): 672-684, 2022 05.
Artículo en Inglés | MEDLINE | ID: mdl-35550615

RESUMEN

The vasculature is an essential organ for the delivery of blood and oxygen to all tissues of the body and is thus relevant to the treatment of ischaemic diseases, injury-induced regeneration and solid tumour growth. Previously, we demonstrated that ETV2 is an essential transcription factor for the development of cardiac, endothelial and haematopoietic lineages. Here we report that ETV2 functions as a pioneer factor that relaxes closed chromatin and regulates endothelial development. By comparing engineered embryonic stem cell differentiation and reprogramming models with multi-omics techniques, we demonstrated that ETV2 was able to bind nucleosomal DNA and recruit BRG1. BRG1 recruitment remodelled chromatin around endothelial genes and helped to maintain an open configuration, resulting in increased H3K27ac deposition. Collectively, these results will serve as a platform for the development of therapeutic initiatives directed towards cardiovascular diseases and solid tumours.


Asunto(s)
Regulación del Desarrollo de la Expresión Génica , Factores de Transcripción , Diferenciación Celular/genética , Cromatina , Nucleosomas , Factores de Transcripción/genética , Factores de Transcripción/metabolismo
20.
Annu Rev Genomics Hum Genet ; 23: 53-71, 2022 08 31.
Artículo en Inglés | MEDLINE | ID: mdl-35440147

RESUMEN

Virtually all cell types have the same DNA, yet each type exhibits its own cell-specific pattern of gene expression. During the brief period of mitosis, the chromosomes exhibit changes in protein composition and modifications, a marked condensation, and a consequent reduction in transcription. Yet as cells exit mitosis, they reactivate their cell-specific programs with high fidelity. Initially, the field focused on the subset of transcription factors that are selectively retained in, and hence bookmark, chromatin in mitosis. However, recent studies show that many transcription factors can be retained in mitotic chromatin and that, surprisingly, such retention can be due to nonspecific chromatin binding. Here, we review the latest studies focusing on low-level transcription via promoters, rather than enhancers, as contributing to mitotic memory, as well as new insights into chromosome structure dynamics, histone modifications, cell cycle signaling, and nuclear envelope proteins that together ensure the fidelity of gene expression through a round of mitosis.


Asunto(s)
Cromatina , Mitosis , Cromatina/genética , Cromosomas/genética , Código de Histonas , Humanos , Mitosis/genética , Factores de Transcripción/genética
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...